Search

Additional file 5: Supplementary Figure S2. Characterization of uncultured umbilical epithelial cells. a. Heatmap showing differentially expressed signature genes in the two epithelial cell subpopulations. b. KEGG and GO analysis of the two epithelial cell subpopulations.

Additional file 3: Supplementary Table S2. List of the top 50 genes differentially expressed in the two MSC subpopulations.

Additional file 1. Methods used in this study.

Additional file 6: Supplementary Figure S3. Expression of fibroblast/stromal marker genes in group 1 and 2 UC-MSCs.

Additional file 4: Supplementary Figure S1. Cultured human UC-MSCs show major MSC features. a. Flow cytometry analysis of cell surface markers on the cultured UC-MSCs. b. In vitro differentiation potentials of human UC-MSCs. Histochemical staining was performed to assess the differentiation into osteoblasts, chondrocytes, and adipocytes. Scale bar: 50 μm.

Additional file 8: Supplementary Figure S5. Expression of skeletal stem cell (A) and pericyte (B) markers genes in the two UC-MSC subpopulations.

Additional file 2: Supplementary Table S1. List of the top 50 genes differentially expressed in the two epithelial subpopulations.

Additional file 9: Supplementary Figure S6. IL17, TGF, and TNF pathway were activated in group 1 UC-MSCs. Genes in red are up-regulated.

Additional file 1: Table S1. Primers for construction of the truncated mutants of HA-fused SH3BGRL. Supplemental Figure S1. SH3BGRL expresses in various breast cancer cell lines. Supplemental Figure S2. SH3BGRL prolongs HER2 stability and duration on cell membrane. Supplemental Figure S3. SH3BGRL promotes cell cycle progression. Supplemental S4. SH3BGRL renders drug resistance. Supplemental S5. Targeting SH3BGRL represses the xenografted tumor formation. Supplemental Figure S6. Targeting SH3BGRL represses cell proliferation and enhances cell apoptosis in xenografted tumors. Supplemental Figure S7. SH3BGRL activates HER2 to promotes tumor progression. Supplemental Figure S8. Relevance of SH3BGRL and HER2 in breast cancer tissues.

Figure S9. Relationship of PCBP1 to p27 mRNA level in tumor samples. (A). Semiquantitative RT-PCR detection of PCBP1, p27 mRNA expression in conlon tumor tissues. GAPDH was used as control. p27 mRNA level is correlated to PCBP1 level in the paired normal (N) and tumor (T) samples of colon tissues. (B). p27 mRNA level as well as its ubiquitin ligase Skp2 in ovarian epithelia. GEO ID is shown. (C). TCGA gene profile of Skp2 in tumors indicates the relative low amplification in ovary and colon tumors. (PPT 2633 kb)

Figure S11. PCBP1 and p27 expression in tumor cells are correlated to anti-cancer drug sensitivity. PCBP1 and p27 expression were analyzed by data from GEO database, and high expression of PCBP1 and p27 were observed in Tamoxifen (GSE26459), Doxorubicin (GSE24460) and Lapatinib (GSE16179) sensitive breast cancer subclones (n indicates the analyzed cell subclone numbers). (PPT 92 kb)

Figure S4. Colonies induced by A2780 or DLD-1 cells overexpressing PCBP1. PCBP1, PCBP1-p27 KD and GFP control cells were analyzed by soft agar assay for anchorage-independent growth. Colonies were stained with Trypan Blue and photographed. (PPT 609 kb)

Figure S7. Expression of PCBP1 and p27 in ovary cancer samples compared to that in the normal tissues. The indicated protein expression level was defined based on the staining intensity under the same robust IHC staining condition. (PPT 1277 kb)

Figure S1. PCBP1 increased p27 mRNA stability in A2780, DLD-1 and MDA-MB-231 cells. (A). Schematic procedure of isolation and identification of PCBP1-bound RNA transcripts. (B). Semi-quantitative RT-PCR detection of p27 mRNA levels in the indicated cell lines. A positive correlation between PCBP1 and p27 mRNA level is observed in these cell lines. (PPT 740 kb)

Figure S10. A positive correlation between PCBP1 and p27 proteinlevels in human renal cancer samples. (B) Relationship between PCBP1 and p27 protein expression in IHC staining. A positive correlation between PCBP1 and p27 protein levels is observed in tumors from 7 patients. (C) RT-PCR detection of PCBP1, p27 mRNA exprission in renal tumor tissues. GAPDH was used as control. (D) Immunoblot of PCBP1 and p27 proteins in healthy tissues (N), tumor adjacent region (A) and tumor tissues (T). GAPDH was used as control. E. Kaplan-Meier survival curve of renal carcinoma patients with different expression level of PCBP1. Data is from https://www.proteinatlas.org/ENSG00000169564-PCBP1/pathology/tissue/renal+cancer . (PPT 2557 kb)

Table S1. Primers for RT-PCR. Table S2. Primers for p27 3â UTR series deletion. Table S3. Primers for PCBP1 mutations. Table S4. Part of mRNAs identified in GFP-PCBP1-RIP only. Table S5. Correlation between clinicopathologic features with PCBP1 expression in renal cancer patients. (DOCX 23 kb)

Figure S5. p27 mRNA stabilized by PCBP1. The half-life of p27 mRNA was derived from the decay curve plotted from the Fig. 6g and h. Linear regression equation in A: PCBP1, y = − 0.0096× + 0.998,R2 = 0.99; GFP, y = − 0.058× + 0.9844, R2 = 0.9865; B: PCBP1 KD, y = − 0.0749× + 0.965, R2 = 0.9607; GFP, y = − 0.0749× + 0.9677, R2 = 0.9387. (PPT 309 kb)

Figure S8. Expression of PCBP1 and p27 in paired colon cancer samples compared to that in the normal tissues. The indicated protein expression level was defined based on the staining intensity under the same robust IHC staining condition. (PPT 1072 kb)

Figure S6. No effect of PCBP2 on p27 expression on both mRNA and protein levels. (A). RT-PCR analysis of PCBP2 knockdown efficiency by 2 specific siRNAs in A2780 GFP control cells or GFP-PCBP1 overexpressing cells. (B). Immunoblot analysis of p27 expression upon PCBP2 knockdown in A2780 GFP control cells or GFP-PCBP1 overexpressing cells. GAPDH was used as a loading control. (PPT 2665 kb)

Supplementary Experimental Procedures. (DOCX 19 kb)

Figure S2. PCBP1 overexpression inhibits cell cycle progression. (A). Representative immunofluorescence staining of A2780 and DLD-1 cells with EdU and Hoechst in cells with endogenous PCBP1 knockdown (PCBP1 KD), GFP-PCBP1 overexpression (PCBP1) and the additional p27KD in PCBP1-overexpressing cells (PCBP1-p27KD). (B). Statistical analyses of EdU positive cells in A. The EdU positive cells were counted randomly in 3 views and analyzed. (mean ± SEM, n = 3 per group). (C). Representative flow cytometry analysis of A2780 and DLD-1 cells with PCBP1 overexpression after EDU staining. Ratios of EdU + GFP-positive cells to GFP positive cells or GFP-PCBP1-positive cells are shown in the right-upper quadrant of each diagram. (PPT 2149 kb)