EMBO/sd-character-level-ner huggingface dataset:
Both FKBDs and the TPR domain of Coi12p are important for scnRNA loading in vivo A A schematic drawing of the in vivo functional analysis of COI12 mutants.Figure 6A A schematic drawing of the in vivo functional analysis of COI12 mutants. The expression constructs from which HA‐tagged Coi12p proteins (wild‐type [WT] or the mutant proteins lacking the two FKBDs [∆FKBD] or the TPR domain [∆TPR]) are expressed under the control of the cadmium‐inducible MTT1 promoter were introduced into the MACs of COI12KO cells. B-G Cells containing the expression constructs shown in (A) were mated with COI12KO cells in the presence of cadmium ions. The localization of Twi1p was analyzed by immunofluorescence microscopy staining using an anti‐Twi1p antibody at 3 hpm (B-D). In (B-D), the parental MACs are marked with arrowheads. DNA elimination was analyzed by DNAFISH with probes against Tlr1 IESs at 36 hpm (E). B-G Cells containing the expression constructs shown in (A) were mated with COI12 KO cells in the presence of cadmium ions. Defects in DNA elimination were categorized as described for Fig C. The expression of the different Coi12p mutants was analyzed by Western blot using an anti‐HA antibody (F, top). Alpha‐tubulin was analyzed as a loading control (F, bottom). The Twi1p‐containing complex was immunoprecipitated with an anti‐Twi1p antibody. Precipitated scnRNA was separated on a denaturing gel and visualized by the nucleic acid staining dye GelRed (G, top and middle). The intensities of bands relative to the sample with the mating expressing wild‐type Coi12p (KO+WT) are shown on the top, and a result of a representative experiment is shown in the middle. Precipitated Twi1p was analyzed by Western blot using an anti‐Twi1p antibody (bottom). In (G), the standard deviation (SD) between technical replicates is indicated.
Figure 4(A) 786-0 cells were transfected with individual siRNAs that targeted 335 E3 ligases (three siRNAs per gene; indicated as 'siGene' on the x axis) for 48 hours and treated with aa starvation for 12 hours in duplicates. Next, the intensity of GDH1-GFP in each well was recorded using Opera Phenix (Perkin Elmer). The data are presented as a volcano plot. The siRNAs with P < 0.0001 (two-tailed Student's t-test) and a GFP intensity ratio (normalized to the non-targeting siRNA) >16 were considered to be effective siRNAs (red). E3 ligases targeted by more than two effective siRNAs were selected as candidates. siE3s, E3 ligase siRNAs; siNC, non-targeting control siRNA. (B) 786-0 cells were transfected with siRNA targeting RNF213, CHFR or TRIM27 for 48 hours and then were subjected to aa starvation for 12 hours. WB was performed using the indicated antibodies. 786-0 cells were treated with MG132 (10 μM) under aa starvation. Antibodies against GDH1 (C) were used to enrich protein complexes in cells with or without GDH1. IgG was used as a control. WB was performed using the indicated antibodies. 786-0 cells were treated with MG132 (10 μM) under aa starvation. Antibodies against RNF213 were used to enrich protein complexes in cells with or without GDH1. IgG was used as a control. WB was performed using the indicated antibodies. (E) 786-0 cells were treated with MG132 (10 μM) under aa starvation. The cells were transfected with siNC or siRNF213 for 48 hours, and then an antibody against GDH1 was used to enrich the GDH1-associated complex. IgG was used as a control. WB was performed using the indicated antibodies. (F) 786-0 cells were transfected with siNC or siRNF213 for 24 hours and then were transfected with or without His-ubiquitin for 36 hours. The indicated cells were treated with or without MG132 (10 μM) under aa starvation. Beads against His were used to enrich His-ubiquitin-associated proteins, which were eluted by PBS containing 250 mM imidazole. WB was performed using the indicated antibodies. (G) Restored expression of WT or 3KR GDH1 in 786-0 cells lacking GDH1 followed by transfection with or without His-ubiquitin for 36 hours. The indicated cells were treated with MG132 (10 μM) under aa starvation. Beads against His were used to enrich His-ubiquitin-associated proteins in the indicated cells. WB was performed using the indicated antibodies. (H) 786-0 or 769-P cells (WT or stably expressing shGDH1) were transfected with siNC or siRNF213 for 48 hours and then were treated with or without aa starvation for the indicated times. Cell viability was determined. Unpaired t-test was used with ***p < 0.001. (I) IHCs of KIRC clinical samples were performed using antibodies against GDH1 or RNF213 (upper panel). The correlated IHC signal between GDH1 and RNF213 in KIRC samples was calculated (lower panel). The cor.test was used to determine the p-value. Scale bars: 50 μm.
Specificity for CLE45 signal transduction resides in the MAKR5 C-terminus.Figure 4(A) Schematic presentation of the MAKR5 protein, indicating the K/R-rich membrane hook motif, the S-rich conserved C-terminus, the position of the makr5 mutation, and the truncated fragments tested. (B) Expression of truncated MAKR5 fusion proteins under control of the MAKR5 promoter in 5-day-old roots (asterisks: protophloem sieve element strands). (C) Close up of protophloem sieve element strands expressing truncated MAKR5-CITRINE fusion proteins under control of the UBQ10 promoter. (D) Failure of truncated MAKR5 fusion proteins to complement CLE45 insensitivity of the makr5 mutant (7-day-old seedlings; 1-2 independent transgenic lines per construct are shown). (E) Sensitivity of makr5 mutant roots to continuous brassinolide (BL) treatment (7-day-old seedlings). (F) Expression of MAKR5-BKI1 fusion proteins under control of the MAKR5 promoter in roots of 5-day-old makr5 seedlings. (G) Complementation of CLE45 insensitivity of makr5 roots by BKI1-MAKR5, but not MAKR5-BKI1 fusion protein (7-day-old seedlings; 3 independent transgenic lines per construct are shown).
Figure 4(A) Timeline of prime-boost LV::S vaccination and SARS-CoV-2 challenge in B6.K18-hACE2IP-THV mice (n = 6/group). The LVs used in this experiment were integrative. (B) Serum neutralization capacity of anti-SCoV-2 Abs in LV::S-vaccinated mice compared to sham mice (n = 6/group). (C) Viral RNA content as determined in various organs at 3 dpi (n = 6/group) by use of conventional E-specific or sub-genomic Esg-specific qRT-PCR. Red lines indicate the qRT-PCR detection limits. Data information: Statistical significance was evaluated by Mann-Whitney test (* = p<0.05, ** = p<0.01). (D) Percentages of NK cells or neutrophils in the lungs of LV::S- or sham-vaccinated and SARS-CoV-2-challenged B6.K18-hACE2IP-THV transgenic mice at 3 dpi (n = 6/group). Percentages were calculated versus total lung live CD45+ cells. Data information: Statistical significance was evaluated by Mann-Whitney test (* = p<0.05, ** = p<0.01). (E) Relative log2 fold change in cytokine and chemokine mRNA expression in the brain of LV::S- or sham-immunized and SARS-CoV-2-challenged B6.K18-hACE2IP-THV transgenic mice at 3 dpi (n = 6/group). Means ± SD are shown. Data were normalized versus untreated controls. Data information: Statistical significance was evaluated by Mann-Whitney test (* = p<0.05, ** = p<0.01).
Lipid overload in autophagy-insufficient state (Atg7+/−) leads to increased accumulation of lipid due to defective lipophagy.f8Lipid overload in autophagy-insufficient state (Atg7+/−) leads to increased accumulation of lipid due to defective lipophagy. Autophagy insufficiency causes delayed turnover of mitochondria and mitochondrial dysfunction, which leads to augmented inflammasome activation when challenged with inflammasome activators such as lipid. Combined effects of increased lipid content, augmented inflammasome activation and their interaction lead to deteriorated insulin resistance and diabetes.
Figure 3A-D PBMCs from vaccine breakthrough cases (n=15) and close contacts (n=26) were examined for T cell responses. PBMCs were left unstimulated (Unstim), were stimulated with pooled SARS-CoV-2 PepTivator® S, S1, M and N peptides for 6 hours (SARS-CoV-2 Peptide Stim), or non-specifically stimulated with phorbol 12-myristate 13-acetate (PMA), then assessed by high-dimensional flow cytometry. CD4+ T cells positive for intracellular staining of IFNγ (a), IL-2 (b), TNF (c), or CD8+ T cells positive for intracellular staining of granzyme B (d) are shown. Error bars indicate median and interquartile range. E To examine polyfunctionality of CD4+ T cells, SARS-CoV-2 peptide-stimulated IFNγ+ CD4+ T cells were further examined for co-expression of IL-2 and/or TNF, and the fraction of cells expressing IFNγ only (1 function), IFNγ and either IL-2 or TNF (2 functions), or IFNγ and both IL-2 and TNF (3 functions) are shown. F The differentiation status of SARS-CoV-2 peptide-stimulated CD4+ T cells (left) and CD8+ T cells (right) were compared based on CD27 and CD45RA expression (Naïve: CD27+ CD45RA+; T central memory (TCM): CD27+ CD45RA-; T effector memory (TEM): CD27- CD45RA-; TEMRA: CD27- CD45RA+). The average of all vaccine breakthrough cases or close contacts are plotted respectively.
Tpl2[D270A] mutation alters the protein composition of phagosomesWT and Tpl2[D270A] BMDMs were incubated with latex beads for 0.Figure 3A. Heatmap of selected proteins that were significantly downregulated in BMDMs from Tpl2[D270A] mice relative to WT (P < 0.05). Selected hits were grouped into clusters according to their molecular functions. B. Gene Set Enrichment Analysis (GSEA) of significantly downregulated biological processes in phagosomal fractions. Changes of Tpl2[D270A] phagosomes relative to WT. Dot colour; enrichment score. Dot size; statistical significance. C. Protein intensities of V-ATPase subunits from phagosomes purified from WT and Tpl2[D270A] BMDMs, (n = 3 biological replicates). Data information: Data were analysed by Student's t-test. Error bars represent SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. D. Protein intensities of RAB5 and LAMP-1 from phagosome proteome analysis of WT and Tpl2[D270A] BMDMs (n = 3 biological replicates) (left). Immunoblot of isolated phagosomes from WT and Tpl2[D270A] BMDMs probed for RAB5, LAMP-1 and vimentin. Phagosomal fractions of two biological replicates were pooled. One representative experiment out of two shown (right) (n = 2). Data information: Data were analysed by Student's t-test. Error bars represent SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
Figure 3Forebrain slices from WT (A-F) and S1928A KI mice (G-L) were treated with vehicle (water) or 10 M isoproterenol (ISO) for 0.5 - 10 min before solubilization, ultracentrifugation, IP of α11.2 (A-C, G-I) or β2AR (D-F, J-L), and sequential IB for pS1928, pS1700, and α11.2, for GluA1, or for β2AR, of corresponding regions of the blots, as indicated. All the α11.2 IPs in A-C and G-I were from the same samples (which were split in half for parallel IP) as the β2AR IPs in D-F and J-L, respectively (for quantification of coIP of β2AR with α11.2 see Figure EV1F,G).(A-F) In WT, the time-dependent increase in S1928 and S1700 phosphorylation (A-C) paralleled the decrease in coIP of β2AR with α11.2 (A bottom, Figure EV1F) and of α11.2 with β2AR (D-F).(G-L) In S1928A KI mice, ISO induced S1700 phosphorylation (G,I) but did not disrupt the α11.2 - β2AR interaction (G bottom, J-L, Figure EV1G).(B,C,H,I) For quantification of α11.2 phosphorylation, pS1928 and pS1700 signals were normalized to α11.2 (**p<0.01, ***p<0.001, ANOVA). Forebrain slices from WT (A-F) and S1928A KI mice (G-L) were treated with vehicle (water) or 10 M isoproterenol (ISO) for 0.5 - 10 min before solubilization, ultracentrifugation, IP of α11.2 (A-C, G-I) or β2AR (D-F, J-L), and sequential IB for pS1928, pS1700, and α11.2, for GluA1, or for β2AR, of corresponding regions of the blots, as indicated. All the α11.2 IPs in A-C and G-I were from the same samples (which were split in half for parallel IP) as the β2AR IPs in D-F and J-L, respectively (for quantification of coIP of β2AR with α11.2 see Figure EV1F,G).(A-F) In WT, the time-dependent increase in S1928 and S1700 phosphorylation (A-C) paralleled the decrease in coIP of β2AR with α11.2 (A bottom, Figure EV1F) and of α11.2 with β2AR (D-F).(G-L) In S1928A KI mice, ISO induced S1700 phosphorylation (G,I) but did not disrupt the α11.2 - β2AR interaction (G bottom, J-L, Figure EV1G).(B,C,H,I) For quantification of α11.2 phosphorylation, pS1928 and pS1700 signals were normalized to α11.2 (**p<0.01, ***p<0.001, ANOVA).(E,F,K,L) For quantification of coIP, α11.2 and GluA1 signals were normalized to β2AR (***p<0.001, One Way ANOVA).
Fig.Figure 1(A) Schematic diagram of NrCAM's domain structure and sequential proteolytic processing. NrCAM is firstly cleaved by furin, then by ADAM10 and finally by the γ-secretase. The red antibody indicates the binding region of the N-terminal NrCAM antibody. ICD (intracellular domain), CTFf (C-terminal fragment created by furin cleavage), CTFα (C-terminal fragment created by ADAM10 cleavage). (B) Detection of soluble, cleaved NrCAM (sNrCAM) and full-length, mature NrCAM (mNrCAM) and soluble APPα (sAPPα) in neuronal supernatants and lysates (prepared from E16 neurons), treated with GI254023x (5 µM), or solvent for 48h. Densitometric quantifications of the Western Blots are shown (** p < 0.01; *** p < 0.001; **** p < 0.0001, two-sided students t-test n = 6-8). (C) Detection of sAPPα, sNrCAM and mNrCAM in ADAM10 cKO neuronal supernatants and lysates at 7 days in vitro (DIV7). The neurons prepared from floxed ADAM10 (ADAM10 fl/fl) mice were infected with a lentivirus encoding improved Cre recombinase (iCre) or a control GFP lentivirus at DIV2. Conditioned media were collected for 48 h. Densitometric quantifications of the Western Blots are shown (** p < 0.01; *** p < 0.001; **** p < 0.0001, two-sided students t-test n = 6-8).
a, Cellular viability of super-repressor- or empty-vector-infected control;Bcl2 lymphomas treated with ADR or untreated for 5 days, then exposed for 2 days to the inhibitors 2DG or bafilomycin A1, compared to no inhibitor (ut) in vitro.f4a, Cellular viability of super-repressor- or empty-vector-infected control;Bcl2 lymphomas treated with ADR or untreated for 5 days, then exposed for 2 days to the inhibitors 2DG or bafilomycin A1, compared to no inhibitor (ut) in vitro. Results represent means ± s.d. (n = 5 each). b, Immunoblot analyses of ATF4, global protein ubiquitination, p62/SQSTM1, and MAP1-LC3-I to MAP1-LC3-II conversion in control;Bcl2 lymphomas with no ADR treatment, in TIS cells, and in TIS cells after exposure to 2DG or bafilomycin A1in vitro; representative blot with α-tubulin as a loading control (gels were cropped to ease presentation; full-length blots are provided in Supplementary Fig. 16). c, Cleavage (activation) of caspase 12 and detection of cleaved caspase 3 by immunoblot analysis of control;Bcl2 lymphomas as in b; representative blot with α-tubulin as a loading control (gels were cropped to ease presentation; full-length blots are provided in Supplementary Fig. 16). d, Cellular viability of ADR-treated control;Bcl2 lymphoma cells infected with short-hairpin RNAs against either caspase 12 (shCasp12) or caspase 3 (shCasp3) compared to empty vector-infected lymphoma cells treated as in a. Results represent means ± s.d. (n = 4 each). e, TIS cells in primary human acute myeloid leukaemia (AML) samples by SA-b-gal staining of treated (0.1 µg ml−1 ADR for 5 days) versus untreated cells in vitro. Data represent means ± s.d. (n = 5 each). f, Intracellular glucose (left) and ATP (right) concentrations in senescent AML cells as in e. g, Cellular viability of primary AML samples treated as in e and exposed to inhibitors as in a. Results represent means ± s.d. (n = 5 each). h, Colour-coded whole-body fluorescence imaging to visualize green fluorescent protein (GFP)-tagged tumour burden in time-courses of three representative mice harbouring CTX-senescent control;Bcl2 lymphomas at day 5 (see Figure 1a), and two days later, after the administration of 2DG (n = 3 mice), bafilomycin A1 (n = 4 mice) or mock treatment (n = 4 mice) starting from day 5 post CTX (red, highest intensities; blue, lowest intensities). TdT-mediated dUTP nick end labelling (assay) (TUNEL) staining of lymph node sections in situ to visualize apoptosis at day 7. Numbers represent means ± s.d. (n = 3). i, Overall survival of 5 day-CTX-treated control;Bcl2 or Suv-39h1-;Bcl2 lymphoma-bearing mice randomly assigned to sequential exposure to bafilomycin A1 or mock as in h (control;Bcl2: n = 12 and Suv39h1-;Bcl2: n = 10), presented as matched pair analyses (P<0.001 for the control; Bcl2 comparison; P = 0.221 for the Suv39h1-;Bcl2 comparison). *P<0.05.
Figure 3U2OS ATG2A/B double knock-out (DKO) CRISPR/Cas9 cells stably expressing Tandem tagged LC3B (mCherry-GFP-LC3B) were retrovirally transduced to express vector, or HA-tagged ATG2A-WT, -mLIR (FCIL/AAAA) or -mYFS (YFS/AAA). Cells were grown in complete medium (CM) or starved for 2 h (EBSS) or treated with CM plus BafilomycinA1 (200nM, 4 h), fixed and analysed by confocal microscopy. Merged images of GFP (green) and mCherry (red) channels show the presence of autophagosomes/phagophores (GFP and mCherry positive, yellow puncta) or autolysosomes (mCherry only, red puncta) Scale bar 10µm. Images are representative of n=3 independent experiments. Quantification using flow cytometry of measuring GFP and mCherry fluorescence. Cells were gated based on GFP and mCherry fluorescence and % mCherry positive cells gated used as an indication of autolysosome formation due to GFP quenching. Each symbol represents 1 independent experiment with 10,000 cells analysed per condition. A total of n=3 independent experiments were performed, and horizontal bar indicates mean ± SD. U2OS ATG2A/B DKO cells reconstituted with vector only, HA-ATG2A-WT, -mLIR and -mYFS were stimulated with complete medium, (CM), 2 h starvation (EBSS) or 4 h BafilomycinA1 (BafA1, 200nM), lysed in total cell lysis buffer and subjected to western blot analysis. Blots were probed for the presence of HA-tag (ATG2A), p62/SQSTM1, LC3B, pan-GABARAP (GABARAP, GABARAP-L1 and GABARAP-L2) and vinculin (loading control). D, E p62/SQSTM1 (D) and LC3B-II (E) levels were normalized to loading control and quantified as fold change of DKO proteins levels. Each symbol represents an independent experiment. Quantification of at least n=3 independent experiments is shown. Horizontal bar represents mean ± SD.
GATA6 is expressed in MITF-low melanoma cells in vivoA.Figure 8A. Tumor sections were immuno-labelled (IHC) with anti-GATA6 (red) and anti-SOX10 (green) antibodies and images were captured by confocal microscopy at the indicated magnification. We analysed six tumor sections of metastases and observed significant GATA6 expression in only one of them. Scale bar 250 μM for 40x, 100μM for 100x. B. Seurat cluster Heatmap was generated from published scRNA-seq performed on PDX tumour (n=674 cells) (GSE116237) The Heatmap shows 9 different clusters into which the cells can be divided according to the expression of different referenced genes (Z-score). The top 10 genes are indicated in the left for each cluster. C. GO was used to analyze the genes characterizing each cluster identified above. The average P-value was retrieved for each cluster taking the 3 best GO per cluster, then z-score ((P-value of each biological process-average of P-value of each biological process)/standard deviation) was calculated. Clusters 2 and 3 were undefined (un) D-E. Graphs showing the average expression of GATA6 (d) and MITF (e) (RPKM) for each phenotype cluster in T0 (drug naïve) (blue), phases 1-2 (MDR) (green and yellow) and phase 3 (drug-resistance) (red). Data information: data are presented as mean values + SEM (n=6,574 cells from 5 PDX).
Figure 1A. Representative immunofluorescence images and quantifications showing that 3 months old Hnf1apKO mice have increased number of KI67+ (red) acinar cell nuclei co-staining with DAPI (blue) and Amylase (green). Arrows point to KI67+ acinar cells in Hnf1apKO mouse. Scale bar is 200 μm. Acinar proliferation is represented as the average of the KI67+/Amylase+ cell ratio. Quantifications were performed on 3 random fields from 3 Pdx1Cre and 3 Hnf1apKO mice. P-values are from two-tailed Student's t test. Representative H&E stainings of pancreata from KrasG12D and Hnf1apKO;KrasG12D mice. B-D. KrasG12D and Hnf1apKO;KrasG12D mice have normal morphology at 7 days. E-J. At 21 days Hnf1apKO;KrasG12D mice show acinar-to-ductal metaplasia (dashed encircled areas) and regions with desmoplastic reaction (asterisk), which are not observed in KrasG12D mice (E, H). K-P. At 8 weeks KrasG12D pancreas show occasional abnormal ductal structures (dashed encircled areas in N which is a magnification of squared dotted box in K) and Hnf1apKO;KrasG12D mice (L,M,O,P) present mucinous tubular complexes (black arrows), and more advanced PanINs with luminal budding (open arrows) including foci of spindle cell proliferation (asterisks) and incipient infiltrative growth (black dashed box area in O). Data information: Black dashed boxes in (E,F, K, L and O) indicate magnified areas in (H, G, N, M and P) respectively. Scale bars indicate 100 µm (C,E,F,K,L), 50 µm (O) and 20 µm (B,D,G,H-J,M,N,P).
Figure 1Trajectory plots of siRNA-transfected HUVECs under VEGF and/or Sema3E gradients for 12 h. Red trajectories indicate cells with displacement to the negative y-axis at the end of analyses. Rose plots of cell trajectories in (A). Leaflets are grouped into 10° segments. The radius of each leaflet represents the cumulative number of cells migrating in the corresponding angular bin. P values for the Rayleigh test represent non-random distributions of cell endpoints. Forward migration index (FMI) along the y- and x-axes during cell migration. n = 90 cells per group. Data represent mean ± SEM. **p < 0.01; ***p < 0.001; NS, not significant, by Tukey-Kramer test.
Figure 3Representative images (A) and quantitative analysis (B) of IF staining and 3D reconstruction on Sox2-positive region for WT vs. Prom1 KO mice. Ant: Anterior; Post: Posterior (A). Statistics was performed using Student's t-test: **, p < 0.01. Data are represented as mean and standard deviation. Representative IF double staining of Sox 2 (green) and Ki67 (red) on the and transit amplifying cell regions of WT vs. Prom1 KO mice on frontal sections. Samples are counterstained with DAPI (blue). Quantification of Ki67+ cell number in Sox2-positive region based on IF data from WT and Prom1 KO mice (C). Illustration of CLE tissue recombination assay. P7 CLE were microdissected then dissociated with Dispase II to achieve epithelium and mesenchyme. The Prom1 KO epithelium then reassociated with WT mesenchyme or vice versa. Stereo images of three explants cultured with indicated recombination conditions before harvesting at day 3. Dotted line indicates the CLE.
Compromising Hsc70 binding potentiates expression of a heat shock reporter in HSF1-/- mouse embryonic fibroblasts (MEF).Figure 6A Schematics of the used system. MEFs, in which HSF1 on chromosome 8 was deleted, were stably transfected with mTag-BFP expressed under the control of the HSPA6 promoter and subsequently transiently transfected with plasmids that expressed HSF1wt or mutant variants in an artificial operon with hrGFP, which is translated from an IRES element. B Exemplary flow cytometry data plotting BFP fluorescence versus GFP fluorescence, indicating the gates used for analysis. Numbers indicate the fraction (%) of cells (forward scatter singlets) in each gate. Color indicates the cell density from dark blue (low) to red (high). C-H Geometric mean of BFP (C, E) and GFP fluorescence (D, F, H) of cells grown at 37°C (non-stress, NS) or incubated for 1 h at 43°C with a recovery period of 3 h at 37°C (heat shocked, HS) in the GFPhigh (C, D), GFPlow (E, F), and GFPlow-BFPhigh (H) gates. (G) Geometric mean of BFP fluorescence weighted by the relative number of cells in the GFPlow-BFPhigh gate (fluorescence * fraction forward scatter singlets). Note that the GFPlow gate also comprises the GFPlow-BFPhigh gate. Hsf1-202SSS, Hsf1-I202S,L203S,V205S; Hsf1-209SSS, Hsf1-I209S,L211S,L213S; Hsf1-427SSSS, Hsf1-M427S,L429S,L432S,L436S; Hsf1-465SSS, Hsf1-L465S,V466S,Y468S; Hsf1-473SSSS, Hsf1-L473S,F474S,L475S,L476S