EMBO/soda-nel huggingface dataset:
HOTAIR lncRNA promotes epithelial-mesenchymal transition by redistributing LSD1 at regulatory chromatin regions (2021)
Figure 4

Figure 4 - HOTAIR expression changes H3K27 acetylation and LSD1 occupancy of gene promoters.

A Venn diagram representing intersections of gene promoters possessing H3K27ac peaks within the 2kb window around their TSS in Epi cells.

B Metagene profiling within the 10 kb window around TSS of H3K27ac, LSD1 and H3K4me2 over IgG in Epi cells expressing none (CTR) or different HOTAIR variants (HOT, HOTΔP and HOTΔL). bp stands for base pairs.

C Heatmap of unsupervised hierarchical clustering based on H3K27ac mean values at gene promoters showing separation of two phenotypic groups.

D Biological process pathways identified by DAVID as enriched for Class 1 or Class 2 genes hyperacetylated in low- or high-migrating cells, respectively (adj. P-value < 0.05 for Class 1 and P-value < 0.05 for Class 2). Dots represent GO term enrichment with color coding: red for high enrichment, blue for low enrichment, and sizes of the dots representing the significance score of each subset (the bigger the dot, the higher its significance).

E Visualization of the H3K27ac, Lsd1 and H3K4me2 CUT & RUN and of RNA-seq tracks of the TGFB2 promoter region in Epi-CTR, -HOT, -HOTΔP and -HOTΔL cells.
HOTAIR lncRNA promotes epithelial-mesenchymal transition by redistributing LSD1 at regulatory chromatin regions (2021)
Figure 5

Figure 5 - HOTAIR expression perturbs LSD1 activity at enhancers.

A Venn diagram representing intersections of active enhancer regions in Epi cells.

B Diagram representing putative active enhancers identified by LILY in each cell line and shared between low- or high-migrating groups respectively.

C Metagene profiling of H3K27ac, LSD1 and H3K4me2 over IgG within the 5 kb window of enhancers in Epi cells expressing none (CTR) or different HOTAIR variants (HOT, HOTΔP and HOTΔL). bp stands for base pairs.
Mammalian CDC14 phosphatases control exit from stemness in pluripotent cells (2022)
Figure 1

Figure 1. Altered brain development in CDC14-deficient mice.

a, Survival curves of wild type (n=27) and Cdc14 DKO (n=51) mice during the first 6 weeks of life. (***, p<0.001; Log-rank, Mantel-Cox test).

b, Representative images of postnatal day 9 Cdc14 DKO and wild-type littermates.

c, Weight of Cdc14 DKO and wild-type littermates during the first 3 weeks of life (n=4 per genotype).

d, Representative macroscopic pictures of brain dorsal view of wild type (WT) and Cdc14 KO mice at postnatal 0, 5 and 9 days (P0, P5 and P9). Scale bar, 2 mm. The bar plots show the quantification of brain width, length and area, respectively.

e, Representative cresyl violet staining images of midsagittal sections of cerebellar vermis of WT and Cdc14 KO mice at P0, P5 and P9. Fissures underdeveloped in mutant samples are labeled with arrowheads. Scale bar, 700 μm. The bar plots show the quantification of the cerebellar vermis area, the number of folia and the average length of the fissures, respectively.

f, Detailed cresyl violet staining images of midsagittal sections of cerebellar vermis of the same samples. The molecular layer (ML), internal granule cell layer (IGL) and external granule cell layer (EGL) are indicated. The width of these layers is shown in the bar plots.

g, Representative images of misdigital sections stained with DAPI (blue), Calbindin (green) and GFAP (red) of cerebellar vermis of wild-type ad Cdc14 DKO mice at 9 days after birth. Scale bar, 50 μm. Plots show the quantification of Purkinje cell dendritic length (average distance of 30 random measurements) and the number of cells per 100 μm from 3 different fields from 3 different mice in each plot.

Data information: Bars indicate mean ± SEM. In a-g, *, p>0.05; **, p<0.01; ***, p<0.001, Student's t-test with Welch's correction. Non-significant data (p>0.05) are not labeled.In d-f, data show the individual data, average and SEM from 3 WT and 3 DKO mice. In f, each point represents the average value of 9 different random measurements across distinctive folia for each mouse.
Mammalian CDC14 phosphatases control exit from stemness in pluripotent cells (2022)
Figure 2

Figure 2. Deficient neural differentiation in CDC14-deficient ESCs.

a. Confocal microscopy analysis of wild-type and Cdc14 DKO embryonic stem cells (ESCs) subjected to neural differentiation. Neuronal marker TUJ1 (green), glial marker GFAP (red) and nuclear marker DAPI (blue) are stained and quantified after 18 days in differentiation media. Each dot represents the ratio between TUJ or GFAP and DAPI area in an image containing at least 100 cells from 4 independent experiments with two different clones. **** p-value<0.0001. Student's t-test. Scale bar 50 μm.

b, qPCR analysis of different neuronal and glial markers in WT and Cdc14 DKO ESCs at day 0 or after 6 and 18 days in differentiation media. Bars indicate the average from 3 biological replicates with technical triplicates per assay. *, p<0.05; Students' t-test.

c, Confocal microscopy analysis of WT and Cdc14 DKO ESC subjected to neural differentiation. Neural progenitor marker Nestin was stained and quantified after 6 days in differentiation media. Each dot represents the ratio between Nestin and DAPI area in an image containing at least 100 cells. ****, p<0.0001; Students' t-test.

d, Heatmap analysis of selected genes obtained from RNA-seq data in the indicated groups. The color scale indicates relative induction (red) or repression (blue)

e, Gene set enrichment analysis (GSEA) of specific pathways during differentiation. NES, Normalized enrichment score.

f, Representative GSEA analysis of pathways down- or up-regulated in Cdc14 DKO cells at day 7 during the differentiation protocol. The Normalized Enrichment Score (NES) value and FDR (q-value) are shown in each plot.
Mammalian CDC14 phosphatases control exit from stemness in pluripotent cells (2022)
Figure 4

Figure 4. Altered phosphorylation of epigenetic regulators in Cdc14 DKO cells.

a. Cdc14 DKO and control (2 clones each) ESCs were subjected to neural differentiation and phospho-residues were analyzed at day 0 (T0) and after 3 days (T3) in differentiation media by mass spectrometry. The Volcano plots show differentially regulated phospopeptides with residues hyper-phosphorylated in Cdc14 DKO cells shown in orange. All other peptides are in grey.

b, List of proteins with the largest number of hyper-phosphorylated [S/T]P sites in Cdc14 DKO cells at day 3.

c, Predicted structure (Jumper et al, 2021) of UTF1 showing the position of the residues hyper-phosphorylated in Cdc14 DKO cells at day 0 (left) and day 3 (right) with the corresponding log(fold change) and p-values when compared to control cultures.

d, Western blot analysis of CDC14B-GFP co-immunoprecipitations. HeLa cells were transfected with UTF1 and GFP or CDC14B-GFP. Cytoplasmic and nuclear soluble proteins were extracted with RIPA, and nucleolar and chromatin-bound proteins were extracted with high salt RIPA. Immunoprecipitations with anti-GFP antibody were performed. Total protein extracts and immunoprecipitates were loaded and UTF (upper panel) and GFP (lower panel) were blotted.

e, Immunoprecipitation of UTF1 upon co-transfection with CDC14B. Total protein was extracted with high-salt RIPA and immunoprecipitations with anti-UTF1 antibody were performed.

f, Expression of active CDC14B, but not a phosphatase-dead (PD) isoform, reduces slow-mobility bands of UTF1. Vinculin was used as a loading control.

g, Quantification of UTF1 phopho-peptides upon UTF1 immunoprecipitation in cells co-transfected with GFP or CDC14B-GFP. Bars indicate the average from two technical replicates (dots).

h, In vitro kinase assay of recombinant human UTF1 with ERK2, CDK1-Cyclin B1, CDK1-Cyclin A2 or in absence of kinases (C).

i, Quantification of recombinant human UTF1 phospho-peptides by mass spectrometry analysis after treatment in vitro with CDK1-Cyclin A2, ERK2 or in absence of kinases (C). Bars indicate the average from two technical replicates (dots).

j, In vitro phosphatase assay of UTF1. Three different fragments of UTF1 (#1, #3 and #4, see Fig EV5c) were phosphorylated by CDK1-Cyclin A2 and subsequently exposed to wild-type or a catalytically inactive form (PD) of CDC14B.
Mammalian CDC14 phosphatases control exit from stemness in pluripotent cells (2022)
Figure 5

Figure 5. Control of UTF1 protein levels by phosphorylation.

a, Quantitative RT-PCR in wild-type (WT) or Cdc14 DKO ESCs of the indicated transcripts during neural differentiation. n=3 independent experiments (±S.E.M.); *, p<0.05; **, p<0.01; ***, p<0.001; Student's t-test.

b, Expression of a CDC14-GFP fusion protein in ESCs before (T0) and after three days (T3) in differentiation conditions. The arrowheads indicate the concentration of CDC14-GFP in the nucleolus before but not after the induction of differentiation. Scale bars, 5 μM. The plot shows the ratio between nucleolar vs. total nuclear URF1 intensity at T0 (n= 26 cells) or T3 (n=31) in cells from two separate experiments. ***, p<0.001, Student's t-test with Welch's correction.

c, Confocal microscopy analysis of UTF1 intensity in WT and Cdc14 DKO ESCs at time 0 (T0) and after 1 day (T1) in differentiation media. The quantification of UTF1 fluorescence mean intensity (FMI) per cell is shown in the plot to the right. Figures and horizontal bars indicate mean ± S.E.M. (n=1269 WT T0 cells, n=2726 WT T1, n=974 Cdc14 DKO T0, n=1173 Cdc14 DKO T1). **, p<0.01; ****, p<0.0001; Student's t-test with Welch's correction.

d, Western blot analysis of UTF1 expression in WT and Cdc14 DKO ESCs at time 0 and after 3 days in differentiation media.

e, Western blot analysis of UTF1 expression in WT and Cdc14 DKO ESCs in absence or presence of the kinase inhibitors roscovitine (R) or trametinib (T).

f, Immunoblot analysis of UTF1 protein stability after transfection of HeLa cells with UTF1 and GFP or CDC14B-GFP. Cells were treated with cycloheximide (CHX) and total proteins were detected at different time points as indicated. Cyclin B1 and β-catenin are used as controls.

g, Western blot analysis of UTF1 WT or mutants in three (UTF1-3KR) or five (UTF1-5KR) lysines in absence or presence of trametinib (T) or roscovitine (R).

h, Immunoblot of UTF1 mutants in SPOP, SIAH or SPOP+SIAH potential binding sites, as well as S48D and S54D phosphorylation sites. Actin was used as a loading control.
Tenascin-C immobilizes infiltrating T lymphocytes through CXCL12 promoting breast cancer progression (2021)
Figure 1

Figure 1 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. 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.

(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.
Tenascin-C immobilizes infiltrating T lymphocytes through CXCL12 promoting breast cancer progression (2021)
Figure 2

Figure 2 Impact of TNC on CD8 TIL in the syngeneic NT193 breast cancer model

(A) Representative staining images (N = 5) of tissue from TNC high (WT/shC) and TNC low (KO/shTNC) tumors (N = 5 per condition) with the indicated antibodies. Arrows point at the respective immune subtype. Scale bar, 50 µm.

(B, C) Quantification of CD8 TIL upon tissue staining as total number per field (B) and in the stroma or tumor cell nest (C), expressed as relative ratio, *** p = 0.0002, Fisher's exact test and in TNC high (N = 5) and TNC low tumors (N = 5). 3 sections per tumor, 8 random fields per section. Mean ± SEM.

(D, E) Apoptosis measurement upon tumor staining for cl. casp3 (D) and signal quantification per field (E) in TNC high and TNC low tumors (N = 7 per condition), 2 sections per tumor, 5 random fields. *** p = 0.0006, Mann-Whitney test, Mean ± SEM. Scale bar, 50 µm.

(F) Representative staining images (N = 5 tumors, 3 sections per tumor) for the indicated molecules. Scale bar, 50µm. Arrows point at CD8 TIL.

(G, H) Gzmb and Perforin expression as determined by qRT-PCR. N = 6 tumors per condition. ** p = 0.0022 for both graphs, Mann Whitney test. Mean ± SEM.

(I, J) Detection (I) and quantification (J) of CD8 TIL in lung metastasis of NeuNT WT and TNCKO mice (N = 3 mice, n = 7 and 12 lung metastases, respectively). Scale bar, 100 µm. ** p = 0.0097, Mann Whitney test. Mean ± SEM.
Tenascin-C immobilizes infiltrating T lymphocytes through CXCL12 promoting breast cancer progression (2021)
Figure 3

Figure 3 CXCL12 expression and binding to TNC

(A) Heat map representation of the most 17 deregulated Panther identified immune related genes in TNC high (WT/shC) and TNC low (KO/shTNC) tumors (N = 2 tumors each).

(B) CXCL12 protein levels in NT193 tumors (ELISA). N = 7 tumors per group, ** p = 0.0035 (WT host, shC versus sh1TNC), ** p = 0.0070 (WT host, shC versus sh2TNC), * p = 0.0379 (TNCKO host, shC versus sh1TNC), * p = 0.0262 (TNCKO host, shC versus sh2TNC), Mann-Whitney test. Mean ± SEM.

(C) Cxcl12 levels in cultured tumor cells (qRT-PCR). N = 5 independent experiments, ** p = 0.0079, Mann Whitney test. Mean ± SEM.

(D) CXCL12 protein levels (ELISA) in conditioned medium (CM) from cultured tumor cells, N = 6 independent experiments, ** p = 0.0022, Mann Whitney test. Mean ± SEM.

(E) Analysis of Cxcl12 (qRT-PCR) in tumor cells upon pretreatment with the indicated inhibitors for 45 minutes (GW, BOP, DMSO and PBS as control respectively) or 60 minutes (Sunitinib) and 6 hours (Cli95) (6 hours DMSO as control) before incubation with TNC for 24 hours. N = 6 independent experiments, ** p = 0.0022, Mann Whitney test. Mean ± SEM.

(F-H) CXCL12 binding to TNC as determined by SPR measurement (F) and negative EM imaging (G), followed by quantification (H). (G) scale bars, 100 nm (left), 50 nm (right), the arrowheads point at gold labeled CXCL12 and the asterisk points at the fibrinogen globe of the murine TNC monomer. (H) Representation of TNC monomer, oligomerization domain (triangle) to form hexamers as seen in (G), FNIII repeats (grey boxes, contant domains, white boxes, alternative domains), fibrinogen like domain (circle). Representative result (3 independent experiments).
Tenascin-C immobilizes infiltrating T lymphocytes through CXCL12 promoting breast cancer progression (2021)
Figure 5

Figure 5 Inhibition of CXCR4 induces infiltration of CD8 leukocytes and macrophages, and reduces tumor cell death and tumor growth (7 weeks model)

(A) Growth of TNC-high (WT/shC) tumors upon cell engraftment and 5 weeks AMD treatment. N = 20 tumors per condition, *** p = 0.0007, unpaired t-test.The central band represents the median, the ends of the box the first and the third quartiles, and the whiskers reach from each quartile to the minimum or maximum.

(B, C) Imaging and signal quantification upon staining for cl. casp3 and DAPI in PBS and AMD treated tumors, N = 5 tumors per condition, 2 sections per tumor, 5 random fields, ** p = 0.0079, Mann-Whitney test. Mean ± SEM. Scale bar, 50µm, arrows point at cl. Casp3.

(D, E) CD8 TIL abundance and localization in PBS and AMD treated tumors, arrow points at CD8 TIL, scale bar, 50µm (D). (E) Quantification of CD8 T cells per randomly chosen fields. N = 5 tumors per condition, 2 sections per tumor, 5 random fields, ** p = 0.0079, Mann-Whitney test. Mean ± SEM.

(F) Gzmb levels in PBS and AMD treated tumors. N = 10 tumors, **** p < 0.0001, unpaired t-test. Mean ± SEM.

(G, H) Heatmap of tumor cell apoptosis (G) and lymphocyte apoptosis (H) associated genes differentially expressed in PBS and AMD treated TNC-high (WT/shC) tumors, N = 2.

(I, J) Abundance and quantification of F4/80+ macrophages in PBS and AMD treated tumors, (I) staining for the indicated molecules, (arrow, macrophages in the stroma; asterisk, macrophages in the tumor nest), (J) quantification of F4/80+ cells inside the tumor nest. N = 5 tumors per condition, 2 sections per tumor, 5 random fields, ** p = 0.0079, Mann-Whitney test. Mean ± SEM. Scale bar, 50 µm.
Tenascin-C immobilizes infiltrating T lymphocytes through CXCL12 promoting breast cancer progression (2021)
Figure 6

Figure 6 Impact of CXCR4 inhibition on lung metastasis

(A) Schematic depiction of the AMD treatment protocol before sacrifice and analysis of the lungs. Intra peritoneal (ip) injection of 5 mg/kg/day AMD into NeuNT mice (daily for 2 weeks, week 27 - 29).

(B) IF tissue staining identifying blood vessel invasions (BVI), surrounded by an EC monolayer (arrow) and expanded EC (asterisk), scale bar, 50µm.

(C) Quantification of BVI with normal (EC monolayer) and aberrant (expanded EC) phenotype expressed as ratio. N = 5 mice (PBS), n = 3 sections, n = 15 BVI, n = 7 PM (parenchymal metastasis); N = 5 mice (AMD), n = 3 sections, n = 12 BVI, n = 2 PM, ****, p < 0.0001, Fisher's exact test.

(D) Schematic depiction of the AMD treatment protocol before sacrifice and analysis of the lungs. Tail vein (iv) tumor cell engrafted mice (WT/shC, WT/shTNC) were ip injected (daily for one week, week 4-5) with AMD 7.5 mg/kg/day. Metastasis assessment in the 5 weeks iv orthotopic grafting model (D-G).

(E) Ratio of mice with and without lung metastasis determined by H&E tissue staining. N = 8 mice (shC, PBS), N = 7 mice (shC, AMD), N = 7 mice (shTNC, PBS), N = 7 mice (shTNC, AMD). p < 0,0001, Chi square test. Fisher's exact test was used to compare the conditions pairwise *, p < 0,05, *** p < 0,001. p = 0,0109 (shC/PBS versus shTNC/PBS), p = 0.0001 (shC/AMD versus shTNC/AMD).

(F, G) Ratio of BVI and PM in lungs from shC (F) and shTNC (G) engrafted mice. N = 8 mice (shC, PBS), n = 4 BVI, n = 13 PM; N = 7 mice (shC, AMD), n = 15 BVI, n = 21 PM; N = 7 mice (shTNC, PBS), n = 8 BVI, n = 9 PM; N = 7 mice (shTNC, AMD). n = 5 BVI, n = 1 PM, *** p < 0,001, **** p < 0,0001. p = 0.0009 (shC/PBS versus shC/AMD) and p = 0.0001 (shTNC/PBS versus shTNC/AMD), Fisher's exact test.

(H) Schematic depiction of the AMD treatment protocol before sacrifice and analysis of the lungs. Mice with mammary gland (MG) engrafted tumor cells (WT/shC) were daily injected peritumorally (pt) for 5 weeks with 5 mg/kg/day (week 2-7). Metastasis assessment in the 7 weeks orthotopic grafting model (H-L).

(I-L) qRT-PCR assessment of Neu (I), Cd8a (J), Ifnγ (K) and Gzmb (L) by qRT-PCR. PBS (N = 16) and AMD (N = 19) treated mice, *, p = 0.0364, Mann whitney test (I), **, p = 0.0086 unpaired t-test (J), **, p = 0.0055, unpaired t-test (K), ***, p = 0.0003 unpaired t-test (L), respectively. The central band represents the median, the ends of the box the first and the third quartiles, and the whiskers reach from each quartile to the minimum or maximum.
Tenascin-C immobilizes infiltrating T lymphocytes through CXCL12 promoting breast cancer progression (2021)
Figure 8

Figure 8 CXCL12 expression in human breast cancer and patient survival

(A) Representative IHC images of CXCL12 intensity and staining pattern in the TMA. N = 219 tumors. Upper panel, scale bar, 400 μm; lower panel, scale bar, 100 μm. Arrows point at punctate staining in the stroma.

(B, C) Histological score of CXCL12 staining in TNC+ and TNC- tumors in combination with CD8+ (above median) and CD8- (below the median). * p = 0.0206, ** p = 0,0040, Unpaired t-test. The central band represents the median, the ends of the box the first and the third quartiles, and the whiskers reach from each quartile to the minimum or maximum.

(D) Kaplan Meier survival analysis of breast cancer patients upon stratification into tumors with a CXCL12 Hscore below or above the median, p = 0.002. Mann-Whitney test.

(E) Survival analysis of breast cancer patients in the GSE 19783-GPL6580 cohort upon stratification into tumors with high or low expression of TNC and CXCL12. Log-rank test. HR and p values are indicated, see also Appendix Fig. S6B.
Long non-coding RNA ELDR enhances oral cancer growth by promoting ILF3-cyclin E1 signaling (2020)
Figure 1

Fig. 1: ELDR is highly expressed in OSCC.

A: Relative expression of ELDR in OSCC patient samples compared to adjacent non-tumor tissues (N=20) analyzed by qRT-PCR. Technical triplicates were used from each sample. 18S rRNA was used as an internal control. Data were analyzed by Student's t test. Small bars indicate standard error (*, p<0.05, **, p<0.01; *** p<0.001).

B: OSCC grade wise upregulation of ELDR expression was noted in the patient samples (N=20). Correlation analysis showed signification positive correlation of ELDR expression with tumor grade (R2= 0.926, significant P value = 0.01). Pearson correlation coefficient and p value was calculated using socscistatistics.com. Well: well differentiated, Mod: moderately differentiated, Poor: poorly differentiated carcinoma. Data are represented as the mean ± SD,

C: Representative images of FISH analysis with an RNA probe of ELDR (red) and DAPI staining (blue) in formalin fixed paraffin embedded OSCC tissue. An unrelated probe was used as negative control. Magnification: 40X; inset magnification 100X and scale bar 10 µm.

D: Relative expression of ELDR in OSCC cell lines compared to normal oral keratinocytes (NOK) analyzed by qRT-PCR. 18S was used as internal control. Data were analyzed by Student's t test. Small bars indicate standard error (**, p<0.01; *** p<0.001). n = 4 biological replicates. Data are represented as the mean ± SD,
Long non-coding RNA ELDR enhances oral cancer growth by promoting ILF3-cyclin E1 signaling (2020)
Figure 2

Fig. 2: ELDR induces OSCC cell growth.

A: Control or ELDR plasmid DNA stably transfected NOKs were analyzed for cell proliferation using Trypan blue exclusion at the indicated time points. Live cell numbers are presented. n = 4 biological replicates. Data are represented as the mean ± SD,

B: Control or ELDR plasmid DNA overexpressed NOK (3×102) were seeded and allowed to form colonies. After two weeks, colonies were stained with crystal violet and counted. Representative images of colonies in control and ELDR overexpressed cells are presented. The right panel shows quantitation. n = 3 biological replicates. Data are represented as the mean ± SD,

C: Cal27, JHU029 and JHU022 cells were transfected with control or ELDR siRNA (50nM), and cell proliferation was analyzed at the indicated time points. n = 3 biological replicates. Data are represented as the mean ± SD,

D: Cell lysates from control or ELDR overexpressing NOKs, control or Cal27 and JHU029 cells treated with two different ELDR siRNAs were subjected to Western blot analysis for PCNA expression using specific antibodies. The membrane was reprobed with an antibody against Actin as internal control for normalization. The bottom panel shows a quantitative representation of Western blot band intensities. n = 3 biological replicates. Data are represented as the mean ± SD,

E: Cal27 and JHU029 cells were transfected with siELDR, ELDR plasmid DNA or both siELDR and ELDR plasmid DNA. Cell lysates were analyzed by Western blot for PCNA expression using specific antibodies. The membrane was reprobed with an antibody against Actin as internal control. The right panel shows quantitative representation of Western blot band intensities. n = 2 biological replicates. Data are represented as the mean ± SD,

Data information: Data were analyzed by Student's t test. Small bars indicate standard error (*, p<0.05; **, p<0.01; *** p<0.001).