EMBO/sd-character-level-ner huggingface dataset:
Figure 8A Immunoblot analysis of protein extracts from siCTRL- or siDDR1/2-transfected SKMEL5 cells plated on FRC- or MAF-derived ECMs in the presence or not of 5 µM BRAFi, 2 µM BRAFi plus 0.01 µM MEKi for 96 h using antibodies against DDR1, DDR2, P-ERK1/2, ERK2, RelB, p100/p52 and HSP60 as loading control were used. B Immunoblot analysis of protein extracts from siCTRL- or siDDR1/2-transfected MM099 short-term cultures plated on FRC- or MAF-derived ECMs in the presence or not of 2 µM BRAFi plus 0.01 µM MEKi for 96 h using antibodies as above. Note that data come from the same immunoblot. C Illustration of the non-canonical p52/RelB NFκB2 pathway and inhibitors used in the study. D Immunoblot analysis of protein extracts from 1205Lu cells cultivated on plastic, Coll-1 or on indicated fibroblast-derived ECMs for 48 h using antibodies against RelB, p100/p52 and ERK2 as loading control. E Immunoblot analysis of protein extracts from 1205Lu (left panels) or SKMEL5 (right panels) cells cultivated on FRC-derived ECMs for 96 h in the presence of 5 µM BRAFi and/or a pan-IKK inhibitor (IKKi, BMS-345541 3 µM) using antibodies against P-ERK1/2, P-Rb, Survivin, RelB, and HSP60 as loading control. F, G Immunoblot analysis of protein extracts obtained from SKMEL5 (F) or MM099 (G) cells plated on FRC- or MAF-derived ECMs and treated with 5 µM BRAFi in combination or not with DDR1-IN-1 (5 µM for SKMEL5 and 3 µM for MM099) or 10 µM NIK inhibitor (NIKi) for 96 h. Antibodies against P-ERK1/2, ERK2, P-Rb, Rb, Survivin, Caspase 3, cleaved Caspase 3, RelB, p100/p52 and HSP60 as loading control were used.
Figure 6A Tumor volumes of eEF2K -/- and eEF2K +/+ RasV12 engraft implanted subcutaneously in AGR129mice. Treatment with NFR or vehicle was initiated 6 days post-implantation. Data are mean of tumor volume ± s.e.m. (n = 8 per group). P values were calculated using 2way ANOVA followed by Bonferroni posttest comparing WT tumor with vehicle or NFR treatment and KO tumors with vehicle or NFR treatment; *P value ≤ 0.05, ** P value ≤ 0.01, *** P value ≤ 0.001. Experiment is representative of 2 performed in same conditions. B Immunoblots for total and phosphorylated eEF2 of tumor engraftment from (A). (NS, Non-Specific signal.)C Bar graph shows relative phosphorylation intensities of eEF2 compared with total eEF2 determined by densitometry analysis. Mean ± s.e.m. are shown (n=4). P values were calculated using 2way ANOVA followed by Bonferroni posttest ; **P value ≤ 0.01.
a, Ter119+CD71+ reticulocytes sorted at day 3 or 6 after PHZ treatment were cultured for in vitro maturation for 0 (top row), 2 (middle row) or 4 (bottom row) days, followed by Mitotracker deep red and Ter119 staining.figf3a, Ter119+CD71+ reticulocytes sorted at day 3 or 6 after PHZ treatment were cultured for in vitro maturation for 0 (top row), 2 (middle row) or 4 (bottom row) days, followed by Mitotracker deep red and Ter119 staining. b, CD71+Ter119+ reticulocytes sorted at day 3 after PHZ treatment were cultured and stained with new methylene blue. Scale bar, 20 µm. c, CD71+Ter119+ reticulocytes sorted at day 3 after PHZ treatment were stained for LC3 and COX IV and analysed by deconvolution microscopy. Scale bar, 5 µm. The Pearson coefficiency for LC3 and COX IV co-localization at day 0 (mean ± s.e.m.) is: wild type, 0.81 ± 0.001; Nix-/-, 0.44 ± 0.036 (n = 35, P = 0.0006). d, CD71+Ter119+ reticulocytes sorted at day 3 after PHZ treatment were analysed by electron microscopy. Scale bar, 0.5 µm.
| Ntras sustains vascular integrity in vivo.Figure 4(A) Validation of Ntras-hnRNPL-interaction by anti-hnRNPL RIPs from H5V lysates, followed by RT-qPCR (n = 4 independent biological replicates). Representative western blot on the right. Data information: In (A, data are represented as mean ± S.E.M. n.s.: non-significant, *p < 0.05, **p < 0.01, ***p < 0.001. (A) two-tailed paired t-test (B) RT-PCR-based analysis of murine Tjp1 exon 20 inclusion in cardiac tissue from control and Ntras-silenced mice (n = 8 mice per group). Experimental outline on the left. Data information: In data are represented as mean ± S.E.M. n.s.: non-significant, *p < 0.05, **p < 0.01, ***p < 0.001. (B, two-tailed unpaired t-test, (C) Illustration of the murine Ntras locus. Displayed are Ntras and Slc30a1 (GRCm38/mm10). The red box highlights the deleted genomic region (~600 bp) in Ntras∆CA/∆CA mice, comprising the hnRNPL binding motif (~130 bp) and flanking sequences (~440 bp). (D) Co-precipitation of Ntras in anti-hnRNPL RIPs using whole heart lysates from NtrasCA/CA and Ntras∆CA/∆CA mice (n = 3-4 mice per group). Representative western blot on the right. Data information: data are represented as mean ± S.E.M. n.s.: non-significant, *p < 0.05, **p < 0.01, ***p < 0.001. two-tailed unpaired t-test, (E) RT-PCR-based analysis of TJP1 exon 20 inclusion in hearts from NtrasCA/CA and Ntras∆CA/∆CA mice (n = 9-14 mice per group). Data information: data are represented as mean ± S.E.M. n.s.: non-significant, *p < 0.05, **p < 0.01, ***p < 0.001. two-tailed unpaired t-test, (F) Retinal angiogenesis assessed in P7 NtrasCA/CA and Ntras∆CA/∆CA pups by immunostaining of isolectin B4. Vascularized areas were normalized to total retinal area (n = 4-8 mice per group). Representative micrographs are shown. Scale bars are 200 µm. Data information: data are represented as mean ± S.E.M. n.s.: non-significant, *p < 0.05, **p < 0.01, ***p < 0.001. two-tailed unpaired t-test, (G) FTSC and TMR-dextran in vivo permeability assays, comparing homogenates of hearts from NtrasCA/CA and Ntras∆CA/∆CA mice. Data normalized to organ and body weight (n = 11-18 mice per group). Data information: data are represented as mean ± S.E.M. n.s.: non-significant, *p < 0.05, **p < 0.01, ***p < 0.001. two-tailed unpaired t-test, (H) Quantification of CD45+ cell (red) infiltration into cardiac tissue from NtrasCA/CA and Ntras∆CA/∆CA mice normalized to DAPI. Isolectin B4 (green) was used to label endothelial cells (n = 4-5 mice per group). Representative micrographs are shown. Scale bars are 50 µm. Insets and arrows indicate sites of CD45+ cell infiltration. Data information: data are represented as mean ± S.E.M. n.s.: non-significant, *p < 0.05, **p < 0.01, ***p < 0.001. two-tailed unpaired t-test, (I) Percentage of control and Ntras-silenced mice showing cardiac pathologies (n = 15-17 mice per group). Prevalence is indicated above the bars. H&E micrographs of murine heart sections from control and Ntras-silenced mice are shown. Insets and arrows indicate sites of lymphocytic infiltration. Scale bars are 100 µm. Data information: (I) Chi-square test.
Figure 7Under the same growth conditions (permitted water, free food, and a 12 h/12 h dark/light cycle at 20±2°C), eight-week-old male C57BL/6 mice were continuously fed with methionine- and choline-supplemented (MCS) or methionine- and choline-deficient (MCD) diets for 6 weeks, and at the same time, gavaged with LicoB, MCC950, or a combination of LicoB and MCC950 (n=8 mice per group (B, C (A) Representative liver section images stained with H&E, Sirius Red, and Masson's trichrome stains. Scale bar: 0.5 cm (top row) and 200 μm (3 bottom ows). (B, C) The serum levels of ALT and AST (n=8 mice per group) were measured using ELISA. Under the same growth conditions (permitted water, free food, and a 12 h/12 h dark/light cycle at 20±2°C), eight-week-old male C57BL/6 mice were continuously fed with methionine- and choline-supplemented (MCS) or methionine- and choline-deficient (MCD) diets for 6 weeks, and at the same time, gavaged with LicoB, MCC950, or a combination of LicoB and MCC950 (n=8 mice per group (D) Western blot analyses of pro-caspase-1, cleaved caspase-1, α-SMA, and ASC in the mice livers, as described in (A). Under the same growth conditions (permitted water, free food, and a 12 h/12 h dark/light cycle at 20±2°C), eight-week-old male C57BL/6 mice were continuously fed with methionine- and choline-supplemented (MCS) or methionine- and choline-deficient (MCD) diets for 6 weeks, and at the same time, gavaged with LicoB, MCC950 n=6 mice per group Real-time quantitative PCR was used to detect the mRNA levels of Col1a1 (E), TNF-α (F) in the mice livers, as described in (A) (n=6 mice per group). Under the same growth conditions (permitted water, free food, and a 12 h/12 h dark/light cycle at 20±2°C), eight-week-old male C57BL/6 mice were continuously fed with methionine- and choline-supplemented (MCS) or methionine- and choline-deficient (MCD) diets for 6 weeks, and at the same time, gavaged with LicoB, MCC950 n=6 mice per group Real-time quantitative PCR was used to detect the mRNA levels of IL-1β (G), and IL-18 (H) in the mice livers, as described in (A) (n=6 mice per group). (I) The levels of IL-18 (I) in the serum was measured using ELISA (n=8 mice per group).
Model for the role of BRCA1‐CtIP in A‐NHEJTelomeres uncapped by TRF2 depletion undergo 3′ overhang excision in MRN‐dependent manner.Figure 7Telomeres uncapped by TRF2 depletion undergo 3′ overhang excision in MRN‐dependent manner. Most telomeres are bound by Ku heterodimer and re‐joined by C‐NHEJ promoted by 53BP1. A subset of the telomeres become substrates for resection reactions, which require MRN, BRCA1 and CtIP. Initiation of resection generates short 3′ telomeric overhangs, whose annealing is facilitated by the homology provided by 2 A‐T base pairs per telomeric repeat. The resulting flaps are cleaved by EXO1 and telomeres re‐joined through the LIG3‐dependent A‐NHEJ pathway.
Figure 5A Enzymatic activity of the succinate dehydrogenase in isolated cardiac mitochondria from 3 animals per genotype. B Mitochondrial membranes were solubilized in 1% digitonin and analyzed by BN-PAGE and Western-blotting using antibodies against SDH components SDHA and SDHB (upper panel). For a loading control, samples were separated on SDS gel and analyzed by Western-blotting (lower panel). C Steady state protein levels analyzed by SDS gel separation of indicated amounts of mitochondrial protein and subsequent Western- blotting with antibodies against SDHA and VDAC3 or visualizing of Flavin fluorescence after excitation with 488 nm. Lower panel: quantification of signal ratio SDHA/VDAC3 in shTAZ and control (set to 100%). D Analysis of gene expression in cardiac tissue analyzed in (C) by qPCR using primers against indicated mRNA. Data of one representative measurement, normalized to b-actin with WT set to 100% are shown. Identical results were obtained in further experiments with two different pairs of animals. E Upper panel: SDH complex of cardiac, liver and kidney were analyzed by BN-PAGE as in (B); lower panel: the same samples were analyzed by SDS gel and Western-blotting. F, G Analysis of enzymatic activity of the succinate dehydrogenase related to the malate dehydrogenase activity in mitochondria isolated from kidney (F) and liver (G). Values of 3 animals per genotype are plotted as percent of wild-type. sh, shTAZ.
Patient FMRP contains a novel nuclear localization signal in the C-terminus.Figure 3Transfected HEK293 cells expressing a GFP-tagged FMRP in which the C-terminus is truncated following the G-ins. mutation site (GFP-FMR1ΔCt). GFP-FMR1ΔCt protein is exclusively cytoplasmic, suggesting that the truncation of the C-terminus alone is not sufficient to explain the localization change observed for the patientFMRP.Transfected HEK293 cells expressing a modified version of GFP-tagged patientFMRP (GFP-FMR1G-ins. [NLSmutated]). The novel amino acid sequence in the C-terminus of patientFMRP [RKRTRRKRTWRRLQRKRRSLPNR] contains stretches of arginines and lysines predicted to function as a nuclear localization signal (based on Expasy protein motif scanner, ScanProsite). Mutating adjacent R and K residues into alanines [RARTRAARTWARLQRAARSLPNR] abolishes the nucleolar localization of the patientFMRP, strongly suggesting that the novel amino acid sequence present in the patientFMRP C-terminus is a functional nuclear localization signal (NLS).
/T465(A) Flag-tagged active (PLK1) and kinase inactive PLK1 (K82M) were co-expressed with HA-EBNA2 wt, docking site mutant (S379A) and phosphorylation mutant (S457A/T465V) and tested for activation of an EBNA2/CBF1 responsive promoter reporter in DG75 cells.Figure 4(A) Flag-tagged active (PLK1) and kinase inactive PLK1 (K82M) were co-expressed with HA-EBNA2 wt, docking site mutant (S379A) and phosphorylation mutant (S457A/T465V) and tested for activation of an EBNA2/CBF1 responsive promoter reporter in DG75 cells. Activation of the reporter firefly luciferase gene is shown as relative response ratio normalized to Renilla luciferase activity and shown relative to EBNA2 wt activity. (B) 30 µg of cellular extracts that were produced for luciferase assays were tested for protein expression using Flag-, HA- and GAPDH-specific antibodies.
Figure 3Structure of RO7019009. ELISA analysis of Aβ species in conditioned medium of HEK293/sw cells treated with RO7019009 or vehicle (DMSO). Aβ levels are shown relative to DMSO vehicle control (n = 4 biological replicates). Dose response curve of RO7019009 in HEK293/sw cells measured using the MSD sandwich-immunoassay (n = 3 biological replicates). Dose response curves for inhibition of Aβ42 generation and Notch cleavage by RO7019009 in different cell lines. Comparison of in vitro potencies for Aβ42 inhibition in HEK293/sw cells, IC50 = 14 nM (n = 3) (as in (C)) with those in H4/sw cells, IC50 = 5 nM (n = 7) and N2a/sw cells, IC50 = 2 nM (n = 6) together with that of Notch cleavage inhibition in a HEK293-based Notch1 luciferase reporter cell line, IC50 > 10 µM (n = 2) (all n numbers represent biological replicates). Left panel: Immunoblot analysis of total Aβ in conditioned media of HEK293/sw cells treated with RO7019009 or vehicle (DMSO). Total APPs levels were analyzed to control for normal APP secretion and equal sample loading. Right panel: Quantification of relative Aβ amounts in (E) (n = 2 biological replicates). Immunoblot analysis of Aβ species in conditioned media of RO7019009 treated HEK293/sw cells after separation by Tris-Bicine urea SDS PAGE; synthetic Aβ peptides were loaded as size markers.
Figure 1A. Detection of Sufu and Skp1 after immunoprecipitation of the indicated Flag-tagged F-Box Proteins (FBPs). An Empty Vector (EV) was used as a negative control. HEK293T cells were treated with MLN4924 (2 M) for 5 hours prior collection. B, C. Immunoprecipitation of endogenous Sufu from PC3 (B) and DAOY cells (C) transfected with either an empty vector (EV) or Myc-tagged Fbxl17. Non-specific rabbit immunoglobulin G (IgG) was used as a negative control. Detection of light chains of immunoglobulin (IgG) was used to assess the amount of IgG used for each immunoprecipitation reaction. Treatment with MLN4924 (2 M) was started 5 hours before cell collection. D. Quantification of Fbxl17 mRNA levels in PC3 and DAOY transfected with a non-targeting siRNA (Control) or two siRNAs to Fbxl17 (1) and (2). Cells were serum starved for 24 hours in serum-reduced medium and treated with SAG (100 nM) for 24 hours prior collection. E. Sufu protein levels in PC3 and DAOY cells treated as in D. GAPDH was used as a loading control. Representative image of three independent experiments is shown. F. Sufu protein levels in PC3 and DAOY cells infected with an empty backbone retrovirus (EV) or a retrovirus expressing Myc-tagged Fbxl17. Representative image from three independent experiments is shown. G. Detection of Sufu protein levels following cycloheximide (CHX) treatment for the indicated hours, and Fbxl17 depletion using two different siRNAs. (Hrs=hours). Representative image of two independent experiments is shown
Depletion of lamin A/C or nesprin 2 causes a defect in docking of Arp2 and myosin Va-mediated preciliary vesicles to the basal body.Figure 7A RPE cells expressing shRNAs specific to luciferase (shLuc.), lamin A/C, or nesprin 2 were grown in the growth medium (+ serum) or serum-starved for 1 h (- serum) and then stained for Arp2 (red), CEP164 (a marker for the mother centriole, green) and acetylated tubulin (white). The representative images are shown. Note that Arp2 was found to localize exclusively at the mother centriole (upper panels) or at the both mother and daughter centrioles (lower panels). Scale bars, 0.5 μm. The percentage of cells with Arp2 localized exclusively at the mother centriole in the total counted cells was measured (n=131-240 cells in each group). B The cells were serum-starved for 48 h and then stained for acetylated tubulin (green), Arp2 (red), and γ-tubulin (white). The representative images are shown. Scale bars, 1 μm (upper panels) and 0.5 μm (lower panels). Note that the cell with Arp2 at the both mother and daughter centrioles fails to form cilium. C The cells were serum-starved for 48 h and then stained for myosin Va (red), acetylated tubulin (green) and γ-tubulin (white). Scale bars, 0.5 μm. The cartoon illustrates that myosin Va-associated vesicles (CV+PCV) specifically accumulate at the mother centriole in most control cells (as in the "one centriole" image). CV, ciliary vesicle. PCV, periciliary vesicle. The percentage of cells with myosin Va exclusively at the mother centriole in the total counted cells was measured (n=160-200 cells in each group).
Loss of the TNC protein in NeuNT tumors impacts gene expression and the immune cell infiltrate(A-C) Display of differentially expressed genes in NeuNT tumors (WT, TNCKO) as volcano plot (A) and heat maps (B, C) representing all (A) and immunity associated genes to be low (B) or high (C) in WT tumors.Figure 1Display of differentially expressed genes in NeuNT tumors (WT, TNCKO) as volcano plot representing all associated genes in WT tumors. N = 3 tumors per genotype. (A) Log2 Fold Change (X-axis) and the negative log10 (P.Value) (Y-axis) is displayed, the blue line indicating p = 0.05 (Y-axis), points above the line, p < 0.05. The dashed red lines indicate the log 2 Fold Change cutoff of -0.25 and 0.25, respectively (X-axis). Green dots represent genes that display both statistical significance (p < 0.05) and fold changes -0.25 and 0.25, respectively. Selected genes regulated by TNC are marked in red (type I interferon response) or pink (Cxcl12) dot. Display of differentially expressed genes in NeuNT tumors (WT, TNCKO) as heat maps (B, C) representing immunity associated genes to be low (B) or high (C) in WT tumors. N = 3 tumors per genotype. (D, E) Infiltration of F4/80+ macrophages determined by flow cytometry. (WT, N = 10, TNCKO, N = 7 tumors), * p = 0.0304, unpaired t-test and iNOS+ macrophages (WT, N = 9, TNCKO, N = 7 tumors) * p = 0.0197, unpaired t-test. Mean ± SEM.
Increased Autophagy in npc1 PCs.Figure 6Increased Autophagy in npc1PCsEM of npc1−/− and wild-type PCs from anterior vermis of 48-d-old mice.(A) Low-power field of a typical npc1PC.(B) Magnification of the red box in (A) showing accumulated multivesicular and multilamellar organelles.(C) Magnification of the blue box in (A) showing a large AV (arrowheads) containing ribosomes and membranes.(D) Another AV (arrowhead) with ER cisternae within the lumen.(E) An npc1PC probably in the late stages of degeneration, with many AVs, some with apparently degrading mitochondria (arrowheads).(F) Low-power field of two wild-type PCs.(G and H) Magnification of the red (G) and blue (H) boxes in (F) showing no AVs and very few multivesicular organelles (arrow).(I) Quantification of area occupied by AVs and other multivesicular/multilamellar organelles expressed as a percentage of total cytoplasmic area for seven wild-type and seven npc1−/−PCs; p = 0.0009 for comparing AV areas and p = 0.0003 for comparing other multivesicular/multilamellar areas, indicating both types of organelles occupy significantly more area in npc1−/−PCs.Bar = 5 μm for (A) and (F); for all others, bar = 1 μm.
Figure 5Mitochondrial respiration (OCR) in control (Ctrl) and MBTPS1-knockout (KO) HaCaT cells was quantified in real time using a Seahorse extracellular flux analyser. Right subpanels: Quantiﬁcation of basal respiration, maximal respiration, and OCR-coupled ATP production in mitochondrial respiration (n = 5 biological replicates). B Quantiﬁcation of relevant metabolites in mitochondrial oxidative phosphorylation (OXPHOS) and glycolysis in Ctrl and MBTPS1-KO HaCaT cells (n = 4 biological replicates). C Nonmitochondrial respiration (ECAR) was quantified in real time using a Seahorse extracellular flux analyser. Quantification of non-glycolytic acidification, glycolysis, glycolytic capacity, and glycolytic reserve in Ctrl and MBTPS1-KO HaCaT cells (n = 4 biological replicates). D MBTPS1 knockout led to a decrease in global ATP production in HaCaT cells. Control and MBTPS1-KO HaCaT cells were cultured in 6-well dishes. A standard curve was generated to calculate the sample ATP concentrations using an ATP Lite Luminescence Assay Kit (n = 4 biological replicates). E MBTPS1 knockout significantly increased the generation of mitochondrial reactive oxygen species (Mito SOX) in HaCaT cells. Left panel: Representative immunofluorescence images of Mito SOX in HaCaT cells. Scale bars: 100 µm. Right panel: The immunofluorescence intensity was quantified to calculate the relative Mito ROS level in HaCaT cells (n = 4 biological replicates). F Schematic representation of OXPHOS and glycolysis upon S1P dysfunction.
PLT3 localises to NBs in Arabidopsis thaliana LRPs and CSCs.Figure 5A-C', PLT3-mV expression driven by the PLT3 endogenous promoter in LRP (A) and primary root SCN (B-C') in plt3 mutant Arabidopsis roots. A, Representative image of PLT3-mV expression (yellow) in an LRP showing the subnuclear localisation to NBs. Transmitted light image in grey. B, B', SCN of an PLT3-mV expressing FM4-64-stained (red) Arabidopsis primary root. The magnification of the CSC layer (B') shows the subnuclear localisation of PLT3 to NBs in a CSC. White arrowhead points at a NB. C, C', SCN of an PLT3-mV expressing Arabidopsis primary root. NBs are visible in the CSC layer in C, also in the transversal view of the CSC layer (C'). Arrowheads in B and C point at the QC (magenta) and CSC (cyan) positions. mV = mVenus; LRP = lateral root primordium; SCN = stem cell niche; NBs = nuclear bodies; CSC = columella stem cell. Scale bars represent 10 µm.
Figure 5 A. Experimental scheme of nascent DNA protection assay. RIF1, rif1∆ and rif1-∆C594 strains (VGY85, CMY6, and CMY7 containing thymidine kinase gene insertions) were arrested with α-factor and released in the medium with 1.13 mM IdU. After 18 min of IdU labelling, IdU was removed by filtration and cells were resuspended in fresh medium with 0.2 M HU and 5 mM thymidine. This thymidine chase was included so that any residual fork extension or do novo origin activation occurring during the HU block would produce unlabelled DNA. After 0, 1, or 1.5 hr, DNA combing was performed and IdU tract lengths analysed  B. Specimen IdU tracts on a DNA fibre. DNA fibre is coloured blue and IdU tracts red. Scale bar is 10 µm (= 20 kb)  C. Degradation of nascent DNA in the absence of Rif1. Plot shows distribution of IdU tract lengths obtained from DNA fibres prepared from cells incubated in the HU block for the time indicated. At least 100 tracts were measured for each condition. Black horizontal bars indicate median values. **, and **** indicate p-values less than 0.01 and 0.0001, respectively, obtained by Mann-Whitney-Wilcoxon test. ns means "not significant"
STED imaging of primary hippocampal neurons fixed with either PFA or glyoxal.Figure 4 Figure 4 STED imaging of primary hippocampal neurons fixed with either PFA or glyoxal. Strong differences in labeling patterns can be observed. The images are brighter and less "spotty" for the glyoxal-fixed samples. Structures such as filaments or organelles are more easily detected. Quantification of the fluorescence signal (fold over background) shows that 16 out of 20 stainings are significantly brighter in glyoxal fixed samples compared to the PFA fixed samples. N = 35 - 132 objects. Scale bar = 6 µm for β-actin and α-tubulin, and 3 µm for the other proteins. ** p < 0.01, *** p < 0.001