EMBO/sd-character-level-ner huggingface dataset:
(C and D) Wild-type primary B cells were treated with vehicle control (-), 5 µg/mL anti-kappa antibody or 1 µM LatA for the indicated time. Cells were lysed and analysed by SDS-PAGE followed by immunoblotting with phospho-Syk (pSyk), phospho-CD19 (pCD19), phospho-Akt (pAkt), phospho-ERK (pERK) and total ERK as loading control. The intensity of phosphorylated proteins, normalized to ERK, was referred to the respective unstimulated sample, set as 1.
BLI of luciferase of representative tumor-bearing mice from all experimental groups at day 19 post therapy initiation.
c, GFP::ATG8 and GFP::ATG8atg1Δ; cells were grown as in a. Cell samples were processed for western blot using anti-GFP antibody. Quantification is presented in Supplementary Fig. 3.
G. Vitamin C-supplemental diet (1%) significantly delays diabetes in Rb∆K7 mice. Shown are two independent experiments with 8 mice per group at indicated time points (left), and 28 and 34 Rb∆K7 mice or 37 and 20 control littermates in each group fed on regular or vitamin C diet, respectively (right). Blood glucose was determined in fasting mice at indicated months (left) or at ~9 months (right). Bars denote mean ±SD. Adjusted P values were calculated using one-way ANOVA, Tukey's multiple comparisons test.
D) Representative microscopy images of living haploid S. cerevisiae cells with Okp1_fl or Okp1_cmΔ, with GFP-tagged kinetochore subunits. Image pixels for images of cells with GFP-fusion proteins of Chl4, Iml3, Mif2, Ctf3, Cnn1, Nkp1, or Nkp2 are 2×2 binned; scale bar: 2 μm. The low GFP signal-intensity that we observe for Chl4-GFP and Iml3-GFP in Okp1_fl cells is consistent with the low GFP signal for Chl4-GFP and Iml3-GFP at centromeres that was previously reported (Joglekar et al, 2008; Lawrimore et al, 2011).
(b) NPCs from three different controls and three different SBMA patients were cultured in the presence of JC-1 dye. Results are presented as a Tukey box plots with box corresponding to first to third quartiles from the median and whiskers corresponding to 1.5 quartiles from the median; n = 3 independent experiments, *P 0.05, t-test, t(4) = 3.983.
(F, G) Flow cytometry profiles representing the levels of MPM-2 (F) and H3S10P (G) in RPE cells in control conditions (Control, black), after treatment with 25 μM P22077 for 4 hrs (USP7i, red) or after treatment with 10 μM RO3306 for 8 hrs followed by incubation with 25 μM P22077 and 10 μM RO3306 for 4 additional hrs (CDK1i+USP7i, blue). DNA content was measured with Propidium Iodide (PI). One representative experiment is shown, out of five.
A) Workflow of the surface analysis performed in SEZ6KO and WT neurons. SEZ6 flox/flox neurons were infected with CRE (SEZ6KO) and GFP (WT) virus at DIV2 and metabolic labeling was started at DIV5. Surface analysis was performed at DIV7.
D. UCSC browser snapshot of the zebrafish vascular endothelial associated lncRNA 2 (veal2) transcript. 5' RACE and 3' RACE data confirmed ends of the veal2 transcript.
The number of TFs shared between metabolic pathways is greater than expected by chance for all combination of pairs of metabolic pathways (Dataset EV4). The size of the nodes corresponds to the number of TFs identified for each pathway. The width of the edge linking two metabolic pathway indicates the number of TFs shared between the two pathways.
B, C, Relative Luciferase Units (RLU; normalized to Renilla luciferase and relative to DNA amount) in 293T cells transfected with DNA constructs carrying the wild-type 3´UTRs from the indicated transcripts (B) or the mutated versions of Dnmt3a and Dnmt3b 3´UTRs, downstream of the luciferase reporter (C). Cells were co-transfected with Renilla luciferase as a control of transfection, and a plasmid expressing GFP or miR-203-GFP. Data are represented as mean ± s.d. (n=3 independent experiments).
D. ChIP for H3K4me3 and H3K36me3 in IGVLR and IGVLUR. Each bar represents the mean of 3 ChIPs for H3K4me3 and IgG and 2 for H3K36me3, each assayed with 3 qPCR repeats (statistical analysis as in Panel A).
(C) HT29 cells were infected with the indicated Shigella strains and incubated for 8 h. Aliquots of cellular supernatants were subjected to cytotoxicity assay. *P<0.05; n.s., not significant (one-way ANOVA).
(f) mTOR immunohistochemistry in brains of individuals with dentatorubral-pallidoluysian atrophy (DRPLA) and spinocerebellar ataxia types 2, 3 and 7 (SCA2, SCA3 and SCA7).
Thymocytes isolated from Ifih1-/- and Cd4cre A1fl/fl Ifih1-/- mice were stained with anti-CD4, anti-CD8, anti-CD69 and anti-MHC-I antibodies and were gated on 4SP thymocytes. Representative images are shown (upper panel). The percentages of CD69+MHC-I- (SM), CD69+MHC-I+ (M1), and CD69-MHC-I- (M2) thymocytes were compared among Ifih1-/- (n = 5), Cd4cre A1fl/fl Ifih1-/- (n = 3) and Cd4cre A1fl/fl (n = 3) mice (lower panel). The same data shown in Fig 2F was used for Cd4cre A1fl/fl mice.
IPs from RNase A-treated S-phase HeLa cell lysate using the ALYREF antibody or IgG (B). Western blotting was performed with indicated antibodies. 2% of input was loaded. * indicates a nonspecific band. The white line delineates the boundary where irrelevant lanes have been removed from the same blots
PC3 cells were treated under indicated conditions for 9 days and subjected to BrdU assays to determine cell cycle distributions. ASNase (0.3 U/ml), TFB-TBOA (5 μM).
Cell viability was measuring using Cell TiterGlo in GSCs #1 and #9 pre-treated for 1 hour with vehicle (DMSO) or QVD (20 μM), and treated for 72 hours more with the indicated doses of MPZ. Data were normalized to the vehicle-treated controls and are presented as the mean + s.e.m of 4 independent experiments.
f, Immunoblot of intestinal epithelial cell scrapings for (p-)IRE1α; after IRE1α; immunoprecipitation (IP). beta-actin, loading control of whole lysates.
C Graph quantification of APPL1-, EEA1- and Rab11- endosomes colocalized with mCitrin-VE-cadherin in sh-Control and sh-PI3KC2β hCMEC/D3 cells. Data are shown as percentage of total count endosomes per cell. Data are expressed as mean ± SEM (n=33-35 cells for APPL1 and EEA1 endosomes and n=16-18 cells for Rab11 endosomes; 3 independent experiments); **p<0.001, Mann-Whitney test or unpaired t-test, as accordance.
E The percentage of CD41+CD42+ cells in CD34+ cells expressing AS-RBM15 Exon 1 were determined by FACS analysis after these cells were grown in pro-MK medium for five days. P value was calculated by t test, p<0.01, (n=3, mean ± SD).
B Hippocampal Gadd45a mRNA levels were significantly upregulated in both Gadd45a-overexpression (q24 = 4.094) and Gadd45aK45E-overexpression groups (q24 = 4.726) (n=9 for all groups). Values shown are mean ± SEM; Tukey´s multiple comparison test: * = p<0.05, ** = p<0.01.
Co-IP assays showing that MYB77 interact with UVR8 in a UV-B-dependent manner in whole seedlings and roots Input: immunoblots showing the levels of UVR8 , and MYB77-TAP in the total protein extract. UVR8 IP: the IP products precipitated by the UVR8 antibody. Total proteins (Input) or IP product were probed, on immunoblots, with an anti-UVR8 or anti-Myc antibody, which detects the TAP tag.
(d) RT-qPCR analysis of TCF20 in TCF20 WT, HET, and KO mice reveals that TCF20 was deleted. n=4 samples. Data information: Error bars represent the means ± S.E.M. Two tailed unpaired T-tests were used to analyze the data, n.s. (no significant difference), p<0.05(*), p<0.01(**), p<0.001(***).
A. EPC2-hTERT-EGFR-p53R175H cells grown in OTC stained for RCP. Arrow indicates invasive region. (scale bar = 100 μm).
E The "in silico oligomer" models were constructed in environments containing 100, 50, and 25 randomly and tightly packed 12-mer 30-nm fiber models (Fig. 3E). The nucleosome concentration was 0.5 mM. The 100-fiber model was drawn using MolScript (Kraulis, 1991). The broken-lined squares show magnified regions.
HuH7 cells were transfected with a control gapmer (ga Ctrl) or with two gapmers targeting EGOT (ga·1αEGOT and ga·2αEGOT). One day later, the cells were infected with a moi 0.3 of HCV for 48h (A to D). Cell supernatants and pellets were collected after infection. RNA was isolated from the cells to evaluate the levels of EGOT (A and E), HCV RNA (B) and GAPDH mRNA, used as a reference to calculate the relative levels of the RNAs. Control HCV RNA has been set to 100% and represents values around 107 GE/μg of total RNA. The experiment was performed at least twice in triplicate (n=6). Average values are shown and error bars indicate standard deviations. Asterisks mark significant differences (*** p<0.001 and **** p<0.0001) obtained with a two-tailed t-test. PFU, plaque-forming units.
E, F Stabilisation of a second copy of DDmyc-PKAc1 leads to the complete block of cell growth indicated by plaque assay (E)
Relative expression of stemness markers in RA-induced differentiation mESCs blocked for 5'-tsRNA function with ASOs. (n = 2) Error bars represent SEM, significance was calculated using unpaired t-test.
E RT-PCR spanning the region from C-terminus of NS5 to CS1 region of 3'UTR for P3 pooled WNV-poly(A) and purified viruses. The primer sequences were described in Materials and methods. Two independent experiments in two replicates were conducted and results of a representative experiment were represented. Values represents mean± sd. Data information: Statistical analysis was performed with two-way ANOVA. n.s. no statistical differences.
(E) GST, GST‐LC3B, GST‐GATE16 and GST‐GABARAP were purified from E. coli and bound to beads. HEK293 cell lysates prepared from cells expressing wild‐type (wt) HA‐ULK1 or the DFP mutant were incubated with the beads, and bound proteins analysed by western blotting.
(D) In SIM images from wild type microglia the sub-autophagosomal localization of NLRP3 was analysed in relation to LC3 and CALCOCO2. LC3 and CALCOCO2 intensities from the center of mass of NLRP3 aggregates were normalised to their maximum within the cluster. While the center of NLRP3 aggregates shows near maximal CALCOCO2 intensity, the relative LC3 intensity there was significantly lower; mean ± SEM, LC3: n = 4, CALCOCO2: n = 3; two-tailed t-test **: p < 0.01.
Scatter plots representing the association of CSF PGRN with temporo-parietal FDG-PET uptake within the subjects of the Alzheimer's continuum group (n = 474). Abbreviations: FDG-PET: Fludeoxyglucose positron emission tomography
(F) The distributions of dwell times of the low FRET states observed in ATP were fit with a single exponential (1539 low FRET states). The inset shows average dwell time and error, defined as the standard error based on the number of traces. (G) As in (F), but with high FRET (1773 high FRET states).
Structures. In 2-5A, ATP and several AMPs (two are shown) are connected by (2'-5') phosphodiester bonds (highlighted in red) by OAS1. In PAR, ADPribose (ADPr) units are connected by (2'-1") glycoside bonds between ribose moieties (highlighted in blue) by PARP1. In PAR-2-5A. PAR can be used by OAS1 as an acceptor for addition of AMP residues to the 2'-hydroxyl of the terminal ADRr, which prevents PAR chain elongation by PARP.
O) Effects of overexpression of a constitutively active form of CDC42 on GSIS from MIN6 cells with ABCA12-deficiency (mean±SEM, n=4 biological replicates, *p=<0.05, Students t-test).
(A) Induction of ycbB under the control of the ParaBAD promoter in BW25113 ΔmepK pHV63(ycbB) was studied by the disk diffusion assay. The clear zone around the disk containing L-arabinose indicates that production of YcbB inhibited growth.
EB formation from WT and RBP-KO ES cell lines. WT ES cell-derived EBs formed cystic structures by day 8 of differentiation; most of RBP-KO ES cell-derived EBs were more compact. Scale bar, 600 μm.
A) Representative western blot of mTORC1 activity measured by phosphorylated p70S6K (T389) from 6 to 72 hours after 20Gy in MRC5 fibroblasts. Data are representative of 3 independent experiments;
(E) HeLa wild-type (WT), Tom1 KO and ATG5 KO cells expressing mycBioID-Tollip and HA-Parkin underwent the same conditions as those above and immunoblotted for the indicated proteins.
(B) Plot showing the first two principal components (PCs) in a PCA of the 60 single-plant transcriptomes. (C) Plot showing the first two PCs in a PCA of the 50 single-plant metabolomes. (D) Plot showing the first two PCs in a PCA of the plant phenomes for the 60 plants that were RNA-sequenced. Data information: The plants in panels (B-D) are numbered according to the numbering in panel (A). Plants belonging to different SNP and DOH subgroups are indicated by different markers and marker colors, respectively, and plants sequenced in the second, smallest batch are circled in panel (B).
(E) Immunofluorescence confocal microscopy staining of LC3B. (I) Control cells treated with PBS and serum, (II) PMA‐treated cells. 1. DAPI: blue, 2. LC3B: red and 3. merge. (III) Percentage of PMN with LC3B aggregates. Control conditions (bar 1: 17.7±2.4%), 3‐MA (bar 2: 3±1.01%), PMA (bar 3: 69±2.1%), PMA and 3‐MA (bar 4: 4±1.9%), rapamycin (bar 5: 56±1.6%). Data are representative of five independent experiments and presented as mean±SD; Student's t‐test, *p<0.01 compared to the control (bar 1) or the indicated comparision (bar 3 vs 4).
B) HT29 cells were infected with Shigella WT, ∆ospC1, ∆ospD3, ∆ospC1∆ospD3, or ∆ospC1∆ospD3/ospC1 (∆ospC1∆ospD3 complemented with ospC1) strains and incubated for 8 h. Cell lysates and aliquots of cellular supernatants were subjected to cytotoxicity assay
E Co-immunoprecipitation (IP) analysis on 66R-derived poly-GP and ubiquitin interaction. Lysates from NSC34 cells transfected with 66R and treated with 10 µM MG132 for 16 h, immunoprecipitated for poly-GP and immunoblotted for ubiquitin and poly-GP.
F Short-term losartan treatment rapidly alleviated inflammation through reduction of Tgf-β signaling. Sections of forepaws as in (E) plus sections of forepaws of C7-hypomorphic mice with moderately affected paws treated with losartan for 4 days were stained for tenascin-C, P-Smad2/3, and Cd11b. There is a clear correlation between the extent of fibrosis, as revealed by tenascin-C staining; Tgf-β signaling, as detected by P-Smad2/3; and inflammation, as indicated by Cd11b+ cells in the C7-hypomorphic forepaws. The 4-day losartan treatment efficiently reduced Tgf-β signaling (P-Smad2/3) and inflammation (Cd11b+) in moderately affected paws, as compared to untreated C7-hypomorphic paws with similar degree of fibrosis. Collectively, the data show that TGF-β-mediated inflammation is a driver of disease progression in RDEB and a major losartan target. Scale bars = 100 μm.G, H Quantification of stainings of moderately affected C7-hypomorphic forepaws with or without a 4-day losartan treatment as in (F). Positively stained cells were quantified after background had been subtracted by applying equal threshold. The values were expressed as positive cells per mm2. Values represent mean ± S.E.M. Data were analyzed with unpaired t-test with Welch's correction. (G) P-Smad2/3 staining, *P = 0.033. (H) Cd11b+ cells, *P = 0.027. n = 3 different mice per group.
E Observation of rosette leaf longevity from the 26-day-old vector control, pER8::CDF4, pER8::CDF4 & nced2, pER8::CDF4 & nced3 and pER8::CDF4 & nced2nced3 transgenic plants after 20 μM estradiol induction for two weeks. Phenotype of detached rosette leaves arranged from oldest to youngest. Scale bars indicate 1.5 cm.
E) Evaluation of the endogenous RB expression using H1299 cell line, under normal conditions, transfecting the cells with an increasing amount of the phosphomimetic mutant MDM2(S395D). F) Evaluation of the endogenous RB as in (E) but using C33 cell line.
D Adding CCL19 in both the upper and lower chambers (CCL19 UL) prevented migration of WT and Arghap45−/− naïve T cells whereas the use of Transwell membrane inserts precoated with ICAM-1 (CCL19 + ICAM-1) did not change the pattern of chemotaxis observed using 'bare' membrane inserts (CCL19). E Chemotaxis of naive CD4+ T cells and B cells from WT, Arghap45−/−, and Arhgap45∆T/∆T mice in response to CCL19. F Chemotaxis of activated CD4+ T cells from WT and Arghap45−/− mice in response to CCL19. Data information: In (A) to (F) each dot represents an individual mouse and as shown CD4+ T cells from 3 to 9 mice have been analyzed for each condition. Mice analyzed on the same day are represented by similar dots (see key in bottom right corner). Mean and SD are shown. *P ≤ 0.033, ** P ≤ 0.002, *** P ≤ 0.001, **** P ≤ 0.0001; A, B, C, D, F multiple unpaired T test; E two-way ANOVA test.
(C) Mobility shift assays monitoring phosphorylation-induced mobility changes of Gis1 (two upper rows) and Rph1 (two lower rows) upon hyperosmotic stress in wild type, igo1∆igo2∆ and Cdc55-depleted cells (right-hand side).
(D and E) In vitro palmitoylation assay for the indicated proteins. NBD-Palm-CoA: (N-[(7-nitro-2-1,3-benzoxadiazol-4-yl)-methyl] amino) palmitoyl-CoA. The recombinant cGAS protein used in the assay was indicated by Coomassie blue staining.
B Basal mitochondrial calcium levels in serum-starved WT and Nox4KO MEFs in the presence or absence of Akti (1 µmol/L) normalized versus WT MEF control. n=6/group with >20 cells imaged per experiment.
Immunoblot detection of PHD2 and ACTIN protein levels from lysates of normoxic cultures (n=4) or B16-F10 tumors [CntlKD (n=5), BNIP3KD (n=7) and ATG5KD (n=6)]. Densitometric quantifications relative to ACTIN levels are shown below each corresponding band. In vitro data was analyzed using a one-sample T-test against shCntl whereas the in vivo data was analyzed using a one-way ANOVA with Tukey's multiple comparisons test.
A) Mitochondrial pH at CV SU e measured in HeLa WT in glycolytic (HGlc) and respiring (Gal) conditions. Where indicated, 5 µg/mL oligomycin was added to inhibit CV activity. B) Mitochondrial pH at CV SU e in IF1-KO cells. Conditions: glycolytic (HGlc) and respiring (Gal) with and without oligomycin. C) Mitochondrial pH at CV SU e in IF1-OE cells. Conditions: glycolytic (HGlc) and respiring (Gal) with and without oligomycin. D)
(C) Immunoblotting of mature brown adipocytes (WT-1) stimulated with insulin (100nM) for 5 min (n=4).
Mice were injected with Ctrl or cGAS-KO MC38 cells and treated with PBS or 5-FU. (B) Pictures of tumors and spleens from a representative experiment. Image panels were cropped from the same picture.
(B) Expression of the Pax5 protein in Pax5Jak2/+ pro-B cells. Nuclear extracts of short-term cultured pro-B cells from 3-week-old Pax5Jak2/+ and Pax5+/+ mice were analyzed by immunoblot analysis with an anti-Pax5 antibody recognizing the N-terminal paired domain. The size of marker proteins is indicated in kilodaltons (kDa) to the left.
C. HyPer7 steady state ratios of the indicated cell lines. HyPer7 probes were targeted to the indicated compartments of the respective cell lines. Cells were grown in glucose-containing medium. In the boxplot, the central band is the median. The lower and upper hinges correspond to the first and third quartiles. The upper whisker extends from the hinge to the largest value no further than 1.5 times the inter-quartile range, whereas the lower whisker extends from the hinge to the smallest value no further than 1.5 times the inter-quartile range.
Hela cells were co-transfected with an RFP-based TDP-NLS reporter and GFP or GA175-GFP. 24 h after transfection, cells were treated with rolipram (30 µM) for 16h. (C) RFP-NLSTDPWT mRNA expression levels were measured by qPCR. n=3 biological replicates. Bar-graphs of mean ± SD. One-way ANOVA with Tukey's multiple comparisons test.
D) (Top) Western blot analysis of CrizR1 cells treated with siScrambled or siRNA for CDK7 or CDK9 for 72h. (Bottom) CrizR1 were treated as above, or with DMSO/alvocidib/THZ1 and cells were stained with Annexin V/PI and analysed by flow cytometry for Annexin V+ cells 72h post-transfection (n=2). Annexin siCDK7 P=0.2, siCDK9 P=0.015, alvocidib P=0.0001, THZ1 P =0.004.
Effect of expression of HA-HuR on Ld infection in mice liver. Scheme for animal experiment done with HA-HuR expression in liver is shown. Also the HA-HuR level was detected in the liver tissue by western blot analysis using anti-HA antibody (F).
ELISA of IL-1β secretion by WT or CYLD KO BMDMs that were either untreated (-) or LPS-primed and then treated with the indicated NLRP3 inducers.
(A, B) HeLa cells infected for 4 h with Listeria WT expressing GFP (WT GFP), Listeria PlcA/B− expressing mCherry (PlcA/B− mCherry), or both Listeria strains at a 1:1 ratio, were analysed by IF using an antibody against NDP52 (A). (B) Quantification of the percentage of cells displaying NDP52+ PlcA/B− mCherry from the experiments described in (A), in cells infected with either PlcA/B− mCherry alone or WT GFP and PlcA/B−mCherry (co‐infection). Values are means±s.e.m. n=3. **P0.01.
B Three pairs of si-TRAF3IP3 together with IFNβ-luciferase reporter were transfected into HEK 293T cells. Twenty-four hours after transfection, the cells were infected with (SeV) or without (mock) Sendai virus, which were harvested six hours post infection to measure the luciferase activity.
GST pull-down analysis of interactions between radiolabeled Myc-Stx16 and WT or LDS-mutant GABARAP (GABARAP-Y49A and -Y49A/F104A). Lower panel shows percentages of Stx16 bound to WT or LDS-mutant GABARAP. Data shown as means ± SEM of precipitated Stx16, n = 3.
CD14+ cells of two convalescent individuals were isolated sequentially at d18, d26 and d56 after confirmation of SARS CoV2 infection by PCR. Following M-CSF differentiation, macrophages were stimulated with LPS (red triangle) or S-Protein (red circles) and IL-1β concentration (pg/ml) was quantified upon incubation with nigericin. Blue dashed line indicates mean IL-1β level of SC-naïve individuals stimulated with S-protein and nigericin.
Analysis of Rad53 phosphorylation in G1 prior to Zeocin addition (G1) and at indicated times. The asterisk indicates a non-specific band detected by the anti-Rad53 antibody.
WT and Mecp2 null MEF cells were starved for 48 h and then treated with the HDAC6 inhibitor tubacin (TUB) (1 μM) for 48 h or with DMSO (VEH) or left untreated (UT). Cells were fixed, stained for anti-acetylated α-tubulin and cilia were counted. The graph shows the percentage of ciliated cells out of three independent experiments (mean ± S.E., *p<0.05, **p<0.01, by two-way ANOVA followed by Bonferroni post hoc test). Two-way ANOVA reported a significant genotype effect (F(1, 10) = 17.94; p<0.01), a significant tubacin treatment (F(2, 10) = 4.425; p<0.05) and a significant interaction (F(2, 10) = 7.266; p<0.05).
Representative auto- (green and violet curves) and cross-correlation (CC, BAX-DRP1, grey curves) curves of DRP1-AF488 and BAX-AF633 measured by FCCS in the membrane of GUVs (B). Dash gray line depict raw data and solid lines correspond to data fitting.
Shown is one representative Western blot from two independent experiments of hepatocytes transfected with siRNA targeting ASK1 (siASK1) or non-targeting siRNA control (siCtrl).
(C) CH12F3-2A cells were transfected with WT Phf5a or its mutants (Phf5aR-MF) along with siRNA against Phf5a. An anti-Flag antibody was used to immunoprecipitate and immunodetect Phf5aR-MF. An anti-Phf5a immunoblot confirmed the knockdown efficiency of the endogenous Phf5a. The antibodies used to detect the co-associated proteins are indicated next to the respective blot.
The fraction of pStat3-positive astrocytes in the peri-plaque region was strongly reduced by the treatment with the Stat3 inhibitor (arrowheads indicate pStat3 signals; scale bars, 100 µm; Mann-Whitney test).
(A) HCT116 p53+/+ or p53-/- cells treated with 50 µM etoposide were assayed by Western blot with antibodies for p53, γH2AX, H3K9Ac, panH3, and GAPDH at times 0, 8, 16, or 24 hr post-treatment.
(A) H1299 cells were co-transfected with the indicated expression vectors (p53, 7.5 ng; HA-Ub, 0.5 ug; His-Xpress-MDM2, 2 ug; RPL11-FLAG, 1 ug; HA-GRWD1-FLAG, 1.5 ug; GFP, 0.04 ug) for 48 h and analyzed by immunoblotting with the indicated antibodies. GFP serves as a control protein to show equal transfection efficiencies.