EMBO/sd-character-level-ner huggingface dataset:
Figure 7(A) Immunoblots for FTMT, frataxin, mitoferrin, HSP60, FTH1 and TfR using the hepatic mitochondrial fraction lysate and whole liver lysate in control (C57BL/6J) mice and STAM mice treated with DFP (0, 0.0375, or 0.075 mg/g body weight) (n=6). The expression of FTMT, frataxin, mitoferrin, FTH1, and TfR were normalized to HSP60 or β-actin, respectively. The central horizontal bar and the error bards indicate mean ± SD. The Tukey's honestly significant test was used for statistical analysis. *: P<0.05, **: P<0.01. (B) The mean number of mitophagosome-like structures/100 μm2 from four randomly selected areas in the livers of DFP (0.075 mg/g body weight)-administered mice were compared between control and FTMT siRNA-treated groups for STAM mice (n=6) and DMBA + HFD mice (n=6). The central horizontal bar and the error bards indicate mean ± SD. Two sample t test was used for statistical analysis. NC: negative control siRNA, *: P<0.05. (C) Frozen liver sections of DFP (0.075 mg/g body weight)-administered mice (STAM mice and DMBA + HFD) with or without FTMT knockdown were stained with dihydroethidium. Red fluorescence indicates the ROS production. Scale bar: 100 μm. siNC: negative control siRNA. (D, E) STAM mice (D) were fed a high-fat diet without DFP treatment or orally administered with 0.075 mg/g body weight of DFP from the age of 4 weeks, injected with FTMT siRNA (10 μl/g body weight) or negative control siRNA through the tail vein every 2 weeks for 8 weeks prior to sacrifice, and followed until 16 weeks of age (n=6). DMBA + HFD mice (E) were fed a high-fat diet, treated with DFP and siRNA in the same manner with STAM mice, and followed until 30 weeks of age (n=6). The number and maximum size of liver tumors were compared among the three groups (control without DFP, negative control siRNA, and FTMT siRNA with DFP). The central horizontal bar and the error bards indicate mean ± SD. The Tukey's honestly significant test was used for statistical analysis. *: P<0.05, **: P<0.01. Representative liver images are shown in the upper panel. Scale bar: 1 cm.
Figure 7(A) Western blotting analysis of RIDs of NCoR1 in left ventricular samples from mice infected with AAV9 GFP or AAV9 NCoR1-RIDs (1939-2453)-Flag. (B) VW/BW of mice 2 weeks after sham operation or AAC. n=7:7:12:12. (C) Representative H&E staining of cross sections of left ventricles. Scale bar: 10μm. (D) Representative WGA staining. Scale bar: 10μm. (E) Quantification of cardiomyocyte size. n=6:6:7:8. (F) qRT-PCR analysis of hypertrophy-related genes in left ventricles. n=8:8:11:11. (G) Representative picrosirius red staining of cross sections of left ventricles. Scale bar: 20μm. (H) Quantification of fibrotic areas. n=5:5:7:7. (I) qRT-PCR analysis of fibrosis-related genes in left ventricles. n=8:8:11:11. (J) Ejection fraction measured before, 2 weeks and 4 weeks after AAC. n=6:6. Two-way ANOVA followed by Bonferoni post-tests was used for statistical analysis. (K) Fractional shortening measured before, 2 weeks and 4 weeks after AAC. n=6:6. Two-way ANOVA followed by Bonferoni post-tests was used for statistical analysis.
Figure 1A. Domain organization of human RADX and RPA1, showing location of OB folds, including three DNA-binding domains (DBDs) in RPA1, and RADX mutants used in this study (see also Fig EV1A). B. Predicted in silico folding of OB folds 2 and 3 of human RADX, modeled by Phyre2, showing similarity to the structure of the region encompassing DBD-A and DBD-B in human RPA1 (Fig 1A). C. RADX interacts with ssDNA. Extracts of U2OS cells stably expressing GFP-RADX WT or mutants were incubated with biotin-coupled ssDNA oligo immobilized on Streptavidin beads, washed extensively and immunoblotted with GFP and RPA1 antibodies. D. ssDNA-bound Streptavidin beads incubated with cell extracts as in (C) were washed with buffer containing indicated salt concentrations prior to immunoblotting. E. Lysates of untransfected HCT116 cells were incubated with immobilized ssDNA or dsDNA probes as in (C) and immunoblotted with RADX and RPA1 antibodies. F. Immunoblot analysis of RADX expression in human cell lines. See also Fig EV1B. G. Representative images from in situ proximity ligation assays (PLAs) in BrdU-labeled U2OS and GFP/GFP-RADX cell lines (Fig EV1E,F) using GFP and BrdU antibodies under native conditions. Scale bar, 10 μm. H. Quantification of data in (F) by quantitative image-based cytometry (QIBC) (n≥3000 cells per condition; data from a representative experiment are shown). See also Fig EV1G. I. Soluble and chromatin-enriched fractions of U2OS cell lines stably expressing GFP-RADX alleles were immunoblotted with indicated antibodies.
Figure 3A. A heatmap showing the effect of single and combinatorial drug predictions by machine learning on indicated cytokine release (left), a zoomed view of top 500 drug predictions (middle), and frequency of Ponatinib occurrence in the top 500 predictions (right). A total of 92,234 single and combinatorial drug predictions were generated from 394 drugs. Predicted cytokine release is expressed as % of DMSO control. B. A heatmap showing experimental validation of model-predicted inhibitor response. All inhibitors were tested at 500nM. Green text indicates an FDA-approved drug. Our model predicted Axitinib to be ineffective, therefore, used as a negative control. C. Comparison of kinase inhibition profile of Ponatinib and Baricitinib. D. A schematic showing ponatinib inhibits multiple kinases involved in the SARS-CoV2-NTD-mediated cytokine signaling.
HsAtg4B is redox regulated during starvation in a similar mechanism to HsAtg4A.Figure 9(A) HeLa cells were transiently transfected with GFP-HsAtg4BWT or GFP‐HsAtg4BC78S. At 24 h post‐transfection, cells were starved for 2.5 h in the presence of 4 mM H2O2 after which they were fixed, permeabilized and incubated with anti‐LC3 polyclonal antibodies. Cells were visualized (upper panel) and quantified (lower panel) as explained in Figure 8. Representative autophagosomes are indicated in the upper panel by arrows. The results presented in the lower panel are the means±s.d. of a total of 100 cells from three separate experiments. (*) indicates significance at P0.001. (B) Left panel: HeLa cells transfected and treated as in (A) were lysed in Ripa buffer and 40 μg of each lysate was loaded to 12% SDS-PAGE and subsequently analyzed with anti‐LC3 antibodies and anti‐GFP antibodies to detect the transfected HsAtg4B and anti‐tubulin antibodies as control. (*) indicates non‐lipidated LC3 and (**) indicates lipidated LC3. Right panel: results from three separate experiments, as detailed in the left panel, were analyzed as explained in Figure 8B.
Cryo-EM reconstructions of the 80S*HCV IRES and 40S*HCV IRES complexes.Figure 1(A) Classical, (B) Rolled, and (C) Rotated subpopulations of the 80S*HCVIRES complex. Left column shows an overview of the final cryo-EM reconstructions with the HCVIRES (pink), 40S subunit (yellow), 60S subunit (blue) and tRNA (green). Middle and right column shows mesh representation of the maps and docked models of the 60S and 40S parts, respectively. 28S-5S-5.8S rRNA (blue), 60S proteins (gold), 18S rRNA (yellow), 40S proteins (dark gray), HCVIRES (pink), tRNA (green). (D) Subpopulations of the 80S*HCVIRES complex containing P-site tRNA. The 80S is in the classical configuration and the head tilt is not present. Left column shows an overview of the final cryo-EM reconstructions with the HCVIRES (pink),40S subunit (yellow),60S subunit (blue) and tRNA (green). Middle and right column shows mesh representation of the maps and docked models of the60S and40S parts, respectively.28S-5S-5.8S rRNA (blue), 60S proteins (gold), 18S rRNA (yellow), 40S proteins (dark gray),HCVIRES (pink), tRNA (green). (E) The cryo-EM map was filtered according to resolution. 40S*HCV IRES. Left column shows an overview of the final cryo-EM reconstructions with the HCV IRES (pink), 40S subunit (yellow), 60S subunit (blue) and tRNA (green). Middle and right column shows mesh representation of the maps and docked models of the 60S and 40S parts, respectively. 28S-5S-5.8S rRNA (blue), 60S proteins (gold), 18S rRNA (yellow), 40S proteins (dark gray), HCV IRES (pink), tRNA (green).
Figure 4(A) Representative images and percentage of HeLa cells with more than 5 H3S10-P foci after transfection with the indicated siRNAs and either pEGFP-C1 (RNH1-) or pEGFP-M27 (RNH1+). Mitotic cells were excluded for the analysis by DAPI staining. More than 300 total cells were considered. Data represent mean ± SEM from three independent experiments. *p<0.05, **p<0.01, ***p<0.001 (one-tailed paired t-test). Black stars denote significant increases whereas red stars denote significant decreases. (B) Representative images and H3K9me2 nuclear signal intensity of HeLa cells transfected with the indicated siRNAs and either pEGFP-C1 (RNH1-) or pEGFP-M27 (RNH1+). At least three experiments were performed. The median of each population in a representative experiment with at least 100 cells per condition is shown. Boxes and whiskers indicate 25-75 and 10-90 percentiles, respectively. *p<0.05, **p<0.01, ***p<0.001 (one-tailed paired t-test). Black stars denote significant increases whereas red stars denote significant decreases.
HERV transcripts originate from activated enhancers.Figure 5Schematic of the experiment. Comparison of RepeatMasker regions annotated "LTR" or "SINE", "LINE", or "SVA" with up-regulated CAGE regions (pVal<0.05) as define in the text based on the Jaccard similarity index. The graph also provides the average and the standard deviation (error bars) of Jaccard indexes calculated for 1000 series of randomly selected CAGE regions. Genes activated in myelin+ patient T cells as compared to myelin+ T cells from healthy controls and annotated in GeneHancer as interacting with a regulatory element containing an LTR sequence and activated 2-fold or more (pVal<0.05) in myelin+ patient T cells as compared to myelin+ T cells from healthy controls: histogram shows the log2 of fold change in read counts within the indicated genes. Error bar corresponds to the FDR as reported by DeSeq2. Screen captures from UCSC genome browser showing regulatory elements containing HERV-derived sequences with transcription correlating with that of a nearby gene annotated as linked in the GeneHancer database.
Starvation triggers accumulation of nucleosides, bases and S7P.Figure 2Starvation triggers accumulation of nucleosides, bases and S7P. Wild‐type S. cerevisiae cells growing in minimal media were switched to minimal media containing no carbon, no nitrogen or no phosphate. After the indicated duration of starvation, the metabolome was quantified by LC‐MS. For media composition, see Supplementary Table S2. For the list of absolute concentration of metabolties, see Supplementary Table S3. Data are shown in heat map format, with each line reflecting the dynamics of a particular compound in a particular culture condition. Metabolite levels of biological duplicates were averaged, normalized to cells growing steadily in glucose (time zero), and the resulting fold changes log2 transformed.
Figure 2In flies with two wild-type lilli alleles (lilli +/+), UAS-Foxn1 activity generates rough-eye (GMR-GAL4), small-eye (ey-GAL4), or white-bristle (hs-GAL4) phenotypes, depending on the GAL4 driver used. These morphological disruptions were prevented when the loss-of-function allele lilli00632 was introduced and the number of wild-type lilli alleles was reduced to one (lilli +/-). Pie charts indicate the percentage of flies in which white bristles were present or completely absent (black bristles only); a total of 212 lilli +/+ and 232 lilli +/- flies were scored, after being raised at 25°C. Scale bar, 100 μm.
Figure 6(A) TCGA cohort of pancreatic cancer patients were divided into two groups according to the median level of ARFGAP1 mRNA expression. Overall survival and disease-free survival were compared between these two groups, as shown in Kaplan-Meier curves. Median survivals (in months), log-rank P values, and hazard ratios (HR) with 95% confidence intervals are indicated. (B) MIA Paca-2 cells were transfected with HA tagged ArfGAP1 or RFP1 (control) for 4 days and then cell numbers were counted. Six replicated wells were counted each group. Columns represent Mean ± SEM. Student's t-test *P = 0.028. (C) Left- Expression levels of ArfGAP1 and phosphorylation of S6K1 (pS6K1) at threonine 389 (mTORC1 activity) were analyzed by immunoblotting. Vinculin was a loading control. Right- MIA Paca-2 cells stably expressing shRNAs against ArfGAP1 or GFP as a control were treated with mTOR inhibitor Torin 1 (250 nM) or vehicle control for 24 hours, and then cell diameters were determined using a particle size counter. The curves show the overall distribution of cell size in each group. Mean of average size ± SD (N = 4): shGFP Vehicle 19.55 ± 0.10, shGFP Torin 1 18.76 ± 0.12, shArfGAP1 Vehicle 20.63 ± 0.16, shArfGAP1 Torin 1 19.05 ± 0.09. Student's t-test P values: shGFP Vehicle vs. shGFP Torin 1 (P = 4.891E-05), shGFP Vehicle vs. shArfGAP1 Vehicle (P = 2.546 E-05), shArfGAP1 Vehicle vs. shArfGAP1 Torin 1 (P = 2.38 E-06), and shGFP1 Torin 1 vs. shArfGAP1 Torin 1 (P = 0.007). (D) MIA Paca-2 cells stably expressing short hairpin RNAs (shRNAs) against ArfGAP1 or GFP (control) were plated and treated with mTOR inhibitor Torin 1 (250 nM) or vehicle control for 48 hours prior to cell counting. Columns represent Mean ± SEM (N = 6). Student's t-test shGFP Vehicle vs. shGFP Torin 1 (P = 2.83 E-07), shGFP Vehicle vs. shArfGAP1 Vehicle (*P = 0.025), shGFP Torin 1 vs, shArfGAP1 Torin 1 (P = 0.139), and shArfGAP1 Vehicle vs. shArfGAP1 Torin 1 (P = 1.22 E-07). "ns" stands for "not significant".
Figure 6A. Western blot of pulldown of AP3 incubated with HIS-6 VPS41WT or VPS41S285P. There is no difference in affinity between VPS41WT and VPS41S285P (See Appendix Fig S10 for MBP-AP3 pulldown input). B. Western blot on cellular or secreted secretogranin II (SgII) of PC12 cells VPS41KO, or VPS41KO transduced with HA-VPS41WT or HA-VPS41S285P lentivirus. Cells were washed and incubated for 30 minutes in Tyrode's solution containing 2.5 mM K+ (basal) or 90 mM K+ (stimulated). No difference in secreted sgII levels was observed between VPS41WT and VPS41S285P. C, D. Cellular (C) and secreted (D) secretogranin II (SgII) were measured by quantitative fluorescence immunoblotting (n=3). VPS41S285P rescues intracellular SgII levels and regulated secretion of SgII to the same extent as VPS41WT.
Loss of MOAP-1 promotes p62-mediated hyperactivation of Nrf2.Figure 7A Increased recruitment of Keap1 to p62 bodies in the MOAP-1 deficient LO2 cells. WT and MOAP-1 KO LO2 cells were subjected to IF analysis with anti-Keap1 (green) and anti-p62 (red) antibodies. Insets in the upper panels are enlarged in the lower panels. Nuclei were counterstained with DAPI (in blue). Scale bar: 5 µm. B MOAP-1 deficiency promotes p62/Keap1 interaction. WT, MOAP-1 KO and p62 KO LO2 cells were subjected to co-IP assay using anti-p62 antibody. TCL, total cell lysates.D Diminished Keap1/Nrf2 interaction in the absence of MOAP-1. WT and MOAP-1 KO LO2 cells were subjected to Duolink PLA using anti-Keap1 and anti-Nrf2 antibodies. Red dots represent Keap1/Nrf2 interaction. Nuclei were counterstained with DAPI (in blue). Scale bar: 10 µm.F Loss of MOAP-1 results in marked elevation of nuclear Nrf2. WT and MOAP-1 KO LO2 cells were subjected to IF analysis with anti-Nrf2 (in green) and anti-p62 (in red) antibodies. Scale bar: 10 µm.H MOAP-1 deficiency promotes activation of Nrf2 signaling. Transcript levels of downstream targets of Nrf2 (i.e. HMOX-1, NQO1, SLC7A11, GCLC and G6PD) in WT and MOAP-1 KO LO2 cells were determined by RT-PCR and normalized to GAPDH. I Elevated transcriptional activity of Nrf2 in the MOAP-1 deficient cells is mediated through p62. Transcript levels of Nrf2 target genes (HMOX-1 and NQO1) in WT, MOAP-1 KO and MOAP-1/p62 DKO LO2 cells were determined by RT-PCR and normalized to GAPDH. J Re-expression of MOAP-1 and MOAP-1-L120E, but not the p62 binding defective mutant of MOAP-1, MOAP-1-M3, inhibits p62/Keap1 interaction in the MOAP-1 deficient cells. MOAP-1 KO LO2 cells were transfected with plasmid encoding Myc-tagged MOAP-1, L120E or M3. The transfected cells were then subjected to co-IP assay with anti-p62 antibody. K Re-introduction of MOAP-1 and MOAP-1-L120E, but not the p62 binding defective MOAP-1-M3, reduces Nrf2 transcriptional activity in the MOAP-1 KO cells. MOAP-1 KO LO2 cells were transfected with Myc-tagged MOAP-1, MOAP-1-L120E or MOAP-1-M3 and ARE-Nrf2-firely and renilla luciferase reporter constructs for overnight and subjected to luciferase assay.
Transcriptional downregulation of ISRE/IRF genes in the intestinal epithelium by zinc part 2.Figure 4(A-B) Expression levels (RT-qPCR data) of Mlkl and Zbp1, from the collection of 48, in ileum biopsy samples, showing the reducing effects of zinc in both GRWT and GRDim mice, and the higher expression in GRDim compared to GRWT (N=4-6/group). RT-qPCR data of Mlkl, Zbp1 in ileum biopsy samples of C57BL/6J mice or different TLR full knockout mice. (N=4/23 per group) RT-qPCR data of Ripk3, in ileum biopsy samples of C57BL/6J mice or different TLR full knockout mice. (N=4/23 per group) RT-qPCR data of Mlkl , in ileum biopsy samples of C57BL/6J mice treated with normal drinking water or 25 mM ZnSO4 in the drinking water for 1 week, either or not pretreated with antibiotics (AB) for three weeks. (N=5 per group) RT-qPCR data of Zbp1, in ileum biopsy samples of C57BL/6J mice treated with normal drinking water or 25 mM ZnSO4 in the drinking water for 1 week, either or not pretreated with antibiotics (AB) for three weeks. (N=5 per group)
is induced by cold exposure in BAT (A-E) BATs were isolated from cold-exposed mice (4℃, 72 h) or control mice (30℃, 72 h), then lysed and subjected to western blotting for analysis of the indicated proteins (A).Figure 1BATs were isolated from cold-exposed mice (4℃, 72 h) or control mice (30℃, 72 h), then lysed and subjected to western blotting for analysis of the indicated proteins (A). Relative protein expression levels, normalized to Actin, are shown in (B). mRNA expression levels of the indicated genes were assayed by real-time PCR and normalized to Actb (C). Mitochondrial DNA copy numbers were determined by real-time PCR and normalized to nuclear DNA standards (D). Relative increase of mitochondrial proteins was normalized to mtDNA copy numbers and plotted (E). (F-G) Representative images showing immunohistochemical detection of FUNDC1 in sections of BATs from mice kept at 30℃ or 4℃ for 72 h. Scale bar, 50 μm. The expression of FUNDC1 was quantified by analyzing and normalizing the average optical density of FUNDC1 staining.
Phosphorylation of CtIP is partially dispensable for MR stimulation in the absence of NBS1.Figure 5Nuclease assays with MR and pCtIP on 3'-end labeled dsDNA, incubated for 2 h.   Quantitation of experiments such as shown in (A). Averages shown; n = 2; error bars, range.   Nuclease assays with MRN, MR and various concentrations of pCtIP (mock-treated) or λCtIP on 5'-end labeled dsDNA, incubated for 2 h. Quantitation of experiments such as shown in (C). Averages shown; n = 3; error bars, SEM. Phosphorylated CtIP was mock-incubated (pCtIP, lane 2) or treated with λ phosphatase (λCtIP, lane 3). The proteins were separated on a 10% polyacrylamide gel and stained with Coomassie brilliant blue. Quantitation of kinetic nuclease assays with MRN and pCtIP (mock-treated) or λCtIP. Averages shown; n = 3; error bars, SEM. Statistical denotations, ** (p < 0.01), two-tailed t-test. Quantitation of nuclease assays as in (F), but with MR complex instead of MRN. Averages shown; n = 3; error bars, SEM. Statistical denotations, ns (p > 0.05), two-tailed t-test. Nuclease assays with MR, MRN, pCtIP or pCtIP T847A, as indicated, on 5'-end labeled dsDNA. Reactions were incubated for 2 h.   Quantitation of experiments such as shown in (H). Averages shown; n = 4; error bars, SEM.
ERK activity responses to single GF pulses.Figure 3A-DSingle GF pulse experiments. Cell‐averaged ERs, population average, and StDev range for n = 10 cells. Pulse application indicated by black bars. 3′ (A, C) and 10′ (B, D) pulse. High (A, B) and low (C, D) GF concentrations. ERationale for measuring amplitude and duration of 1st ERK activity peak in response to GF stimulation. Peak amplitude was measured as the ER change from when the GF stimulation starts until the highest ERK activity before adaptation occurs. Peak duration was estimated as the time between the first point before reaching the half‐maximum of the peak in the ascending phase, and the first point after the half‐maximum in the descending phase. FAmplitude of 1st ERK activity peak in response to different sustained or pulsed GF stimulation experiments. Notched boxplots of 1st ERK activity peak amplitude with median, interquartile range (box), and data within 1.5 IQR range of the lower and upper quartiles (whiskers) are shown (n = 20 cells per experiment). GDuration of 1st ERK activity peak in response to sustained and pulsed GF stimulation. Boxplots of 1st ERK activity peak duration with median, interquartile range (box), and data within 1.5 IQR range of the lower and upper quartiles (whiskers) are shown (n = 20 cells per experiment).
Figure 2(A) An assembly of IgG (first row) and DVL3 (second row) ChIP-Seq data in MDA-MB-468 for the ARAP1, RFX5-GBAT2, WDR74, POU2F3, SNORD3D and HLA-A genes, visualized by IGV. (B) An assembly of IgG (first row) and DVL3 (second row) ChIP-Seq data in MCF7 for the ARAP1, RFX5-GBAT2, ZNF672, SSC5D, HLA-A and HLA-E genes, visualized by IGV. (C) ChIP-qPCRs at ARAP1, RFX5-GBAT2, WDR74 and HAUS5 promoters for IgG and DVL3 in MDA-MB-468 (NTC, shA DVL3, and shB DVL3). (D) ChIP-qPCRs at RFX5-GBAT2, COX14, HLA-E and PSMB8 promoters for IgG and DVL3 in MCF7 (NTC, shA DVL3, and shB DVL3). (E) RT-qPCR-based analysis of expression changes of RFX5, GBAT2, WDR74, HCG15, POU2F3, SNORD3B, APOC1P1, HLA-A, TAP1, PSMD8, PDG1 and JUNB genes in MDA-MB-468 (NTC, shA DVL3, and shB DVL3) cells. (F) RT-qPCR-based analysis of expression changes of RFX5, GBAT2, PCAT6, ZNF672, SSC5D, APOC1P1, HLA-A, HLA-F and TAP1 genes in MCF7 (NTC, shA DVL3, and shB DVL3) cells. D
Figure 2Schematic representation of cellular fractionation to generate non-nucleosomal and nucleosomal fractions. WCE: whole cell extract, NF1: nuclear fraction 1, NF2: nuclear fraction 2. Representative immunoblot analysis of NASP, UBR7, and ASF1 distribution in cellular fractions. Relative loading factors are marked e.g. 1x, 0.2x, etc. Numbers below blot indicate UBR7 immunoblot signal relative to 1x loading of non-nucleosomal fraction. Enrichment of non-nucleosomal:nucleosomal UBR7 as estimated from 5x, 10x, and 20x non-nucleosomal loading conditions in Fig 2B (see Fig EV2). Error bars represent mean +/- standard deviation. A two-tailed one sample t-test was used to calculate p-value (italics) associated with mean difference from a hypothetical value of 1.0 (represented by dashed line). Immunoblot of anti-FLAG IP of 3x-FLAG tagged GFP, or FLAG-HA tagged H3.1 or H2B, from the non-nucleosomal fraction of 293T cells. Representative images of HeLa cells transfected with UBR7-FL. DNA is visualized via DAPI staining shown in magenta and immunofluorescence for 3xFLAG is shown in green. The area within yellow dotted box in (E) is magnified 4x in (E'). Scale bar = 5 μm.   Quantification of the nuclear anti-FLAG immunofluorescence intensity in (E). Error bars represent mean +/- standard deviation. One way ANOVA was used to calculate p-values (italics) associated with differences between treatments. Experiment in (E-F) was performed twice.