Fig 1: BMI1 induces extensive expansion of erythroblasts from healthy donor PBMCs(A) A scheme for BMI1 transduction of PBMC-derived erythroblasts, extensive expansion, and terminal differentiation in culture. Erythroblasts established at an early stage (E8) were used for BMI transduction (counted as day 0). Erythroblasts were expanded continuously for at least 60 days and tested for their potential for terminal maturation. SFEM, serum-free expansion medium. (B) Western blot to assess BMI1 protein levels after GFP or BMI1 transgene transduction. (C) Expansion curves of erythroblasts after BMI1 gene transduction. Established early erythroblasts from health donors P152 and 154 were used. (D) Histochemical (Gimesa) staining of BMI1-transduced erythroblasts at different time points. Scale bar, 20 µm. (E) Flow cytometric analysis of surface markers on the BMI1-induced extensively expanded erythroblasts (BMI1-E3) 60 days after BMI1 transduction. (F) A representative normal karyotype of the BMI1-E3 cells 60 days after expansion from donor P152. (G) Flow analysis of surface markers after terminal maturation. (H) Histochemical (Gimesa) staining of erythrocytes after terminal maturation. Scale bar, 20 µm. (I) Flow cytometric analysis of HBB and HBG in erythrocytes after terminal maturation. (J) Quantitative analyses of differentiated erythrocytes expressing HBB and HBG. (K) The levels of enucleation of BMI1-E3 cells at different time points, all after terminal differentiation for 8 days. (L) Quantification of enucleation rate at different time points. (M) Expansion fold of differentiating E3 cells in terminal maturation.
Fig 2: Sorafenib resistance regulated by GINS1 in HCC cells. (A) gene expression of GINS1 in HepG2 sorafenib-resistance and parent cell lines from GSE140202 (GEO database). (B,C) correlation of sorafenib IC50 and GINS1 expression in 17 hepatocellular carcinoma and colon cancer cell lines obtained from CCLE database (B), and the IC50 value of HCC cell lines (C). (D) HepG2 (left panel) and Huh7 (right panel) cells, w/o GINS1 knockdown, were treated with different doses of sorafenib as indicted and the cell viability was assessed by the MTT assay. (E) HepG2 shGINS1 and Huh7 shGINS1 cells, rescued with HRAS or BMI1 overexpression, treated with different doses of sorafenib as indicted and the cell viability was assessed by the MTT assay. Data are presented as mean ± SD.
Fig 3: HFMSCs promoted the expression of PCNA, Bmi1 and SOX9 in the small intestine. (A–I) Expression of PCNA (A–C), Bmi1 (D–F) and SOX9 (H–J) in the sham group (n = 5), IR group (n = 5) and IR + HFMSCs group (n = 5). (J–L) Statistical comparison of MODs of the AOIs of PCNA (J), Bmi1 (K) and SOX9 (L) in the sham group, IR group and IR + HFMSCs group. The expression levels of PCNA, SOX9 and Bmi1 in the IR + HFMSCs group were the highest. Data are presented as the mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).
Fig 4: Elevated BMI1 in GC-chemoresistant bladder cancer conferred poor prognosis. (A) BMI1 mRNA expression in bladder cancer tissues of patients partial response to chemotherapy versus patients complete response to chemotherapy from TCGA-BLCA database. (B) Relapse-free survival of patients in TCGA-BLCA dataset with low versus high levels of BMI1 mRNA. (C) IHC analysis of BMI1 protein expression in bladder cancer tissues of patients resistant to GC chemotherapy and that sensitive to GC chemotherapy, magnification, ×200 & ×400. (D) Statistical quantification of the IHC score of BMI1 staining in bladder cancer specimens from patients resistant versus sensitive to GC chemotherapy. (E) Relapse-free survival of patients with bladder cancer with low versus high BMI1 expression. (F) Progression-free survival of patients with bladder cancer with low versus high BMI1 expression. *P < 0.05. GC: Cisplatin and Gemcitabine; TCGA: The Cancer Genome Atlas; BLCA: Bladder Urothelial Carcinoma; IHC: Immunohistochemistry; TPM: Transcripts Per Million.
Fig 5: Patients with low expression of Hedgehog/EMT markers together with MAP1LC3B upregulation had a better overall survival and were more responsive to platinum therapy. We compared the in vitro responsiveness of cancer cells to oxaliplatin with the clinical outcomes of the ovarian cancer patients by interrogating the TCGA bioportal (ovarian serous cystadenocarcinoma dataset, TCGA Nature 2011). Those patients for whom therapy response status was not available are classified in N/A group. (A) Box plot showing the distribution of MAP1LC3B mRNA expression according to GLI1 mRNA levels (high vs. low group). (B) Kaplan–Meier plot representing the overall survival status of patients stratified on the basis of the differential expression of GLI1 and MAP1LC3B (high GLI1/low MAP1LC3B vs. low GLI1/high MAP1LC3B). (C) Graph showing the response to platinum therapy based on the differential mRNA expression of GLI1 and MAP1LC3B. The histograms report the number of patients that were resistant, sensitive or developed chemoresistance as soon as the chemotherapy started (too-early group). (D) Box plot showing the distribution of MAP1LC3B mRNA expression according to BMI1 mRNA levels (high vs. low group). (E) Kaplan–Meier plot representing the overall survival status of patients stratified in high BMI1/low MAP1LC3B vs. low BMI1/high MAP1LC3B. (F) Graph showing the response to platinum therapy based on the differential mRNA expression of BMI1 and MAP1LC3B. (G) Box plot showing the distribution of MAP1LC3B mRNA expression according to SNAI1 mRNA levels (high vs. low group). (H) Kaplan–Meier plot representing the overall survival status of patients stratified in high SNAI1/low MAP1LC3B vs. low SNAI1/high MAP1LC3B. (I) Graph showing the response to platinum therapy based on the differential mRNA expression of SNAI1 and MAP1LC3B. (J) Box plot showing the distribution of MAP1LC3B mRNA expression according to TWIST1 mRNA levels (high vs. low group). (K) Kaplan–Meier plot representing the overall survival status of patients stratified in high TWIST1/low MAP1LC3B vs. low TWIST1/high MAP1LC3B. (L) Graph showing the response to platinum therapy based on the differential mRNA expression of TWIST1 and MAP1LC3B.
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