Journal: Scientific Reports

Article Title: CBX7 silencing promoted liver regeneration by interacting with BMI1 and activating the Nrf2/ARE signaling pathway

doi: 10.1038/s41598-024-58248-8

Figure Lengend Snippet: CBX7 regulated hepatocyte proliferation, cell cycle, and apoptosis. ( A ) CBX7 expression in liver tissues of PH mice at different time points was tested by RT-qPCR. ( B ) The RNA expression of CBX7 in NCTC 1469 and BNL CL.2 cells was assayed by RT-qPCR. ( C , D ) The protein expression of CBX7 in NCTC 1469 and BNL CL.2 cells was determined by Western Blot analysis. β-actin was a loading control. ( E ) The proliferation of NCTC 1469 and BNL CL.2 cells was evaluated by CCK-8 assay. ( F – H ) Cell cycle of NCTC 1469 and BNL CL.2 were observed by flow cytometry. ( I – K ) Western blot was used to assay CyclinD1 and CyclinE expression in NCTC 1469 and BNL CL.2 cells. ( L , M ) Cell apoptosis was tested by flow cytometry. Values are means ± SD, ** P < 0.01 vs normal tissue. *P < 0.05, **P < 0.01 vs siRNA-NC; # P < 0.05, ## P < 0.01 vs pcDNA-NC.

Article Snippet: Mouse normal hepatocytes NCTC 1469 (CL-0407), were obtained from ProCell (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, RNA Expression, Western Blot, CCK-8 Assay, Flow Cytometry

Journal: Scientific Reports

Article Title: CBX7 silencing promoted liver regeneration by interacting with BMI1 and activating the Nrf2/ARE signaling pathway

doi: 10.1038/s41598-024-58248-8

Figure Lengend Snippet: CBX7 interacted with BMI1 and inhibited BMI1 expression in hepatocytes. ( A ) proteins that interact with CBX7 are predicted by STRING ( https://string-db.org/cgi/input.pl ). ( B ) Co-IP assay was used to verify the mutual binding of CBX7 and BMI1. ( C ) BMI1 expression in liver tissues of PH mice at different time points was tested by RT-qPCR. BMI1 protein and RNA expression in NCTC 1469 and BNL CL.2 cells were analyzed by RT-qPCR ( D , G ) and western blot ( E , F , H , I ), respectively. CBX7 protein and RNA expression in NCTC 1469 and BNL CL.2 cells was detected by RT-qPCR ( J ) and Western blot ( K , L ), respectively. β-actin was a loading control. Values are means ± SD, ** P < 0.01 vs normal tissue. *P < 0.05, **P < 0.01 vs siRNA-NC; # P < 0.05, ## P < 0.01 vs pcDNA-NC.

Article Snippet: Mouse normal hepatocytes NCTC 1469 (CL-0407), were obtained from ProCell (Wuhan, China).

Techniques: Expressing, Co-Immunoprecipitation Assay, Binding Assay, Quantitative RT-PCR, RNA Expression, Western Blot

Journal: Scientific Reports

Article Title: CBX7 silencing promoted liver regeneration by interacting with BMI1 and activating the Nrf2/ARE signaling pathway

doi: 10.1038/s41598-024-58248-8

Figure Lengend Snippet: Silencing BMI1 aggregated the inhibitory effect of CBX7 overexpression on hepatocyte viability and the promotion of apoptosis. ( A ) CBX7 RNA expression in NCTC 1469 and BNL CL.2 cells was detected by RT-qPCR. ( B ) The proliferation of NCTC 1469 and BNL CL.2 cells was evaluated by CCK-8 assay. ( C – E ) Cell cycle of NCTC 1469 and BNL CL.2 were observed by flow cytometry. ( F – H ) Western blot was used to assay CyclinD1 and CyclinE expression in NCTC 1469 and BNL CL.2 cells. ( I , J ) Cell apoptosis was tested by flow cytometry. Values are means ± SD, *P < 0.05, **P < 0.01 vs pcDNA-NC; # P < 0.05, ## P < 0.01 vs pcDNA-CBX7 + si-NC.

Article Snippet: Mouse normal hepatocytes NCTC 1469 (CL-0407), were obtained from ProCell (Wuhan, China).

Techniques: Over Expression, RNA Expression, Quantitative RT-PCR, CCK-8 Assay, Flow Cytometry, Western Blot, Expressing

Journal: Scientific Reports

Article Title: CBX7 silencing promoted liver regeneration by interacting with BMI1 and activating the Nrf2/ARE signaling pathway

doi: 10.1038/s41598-024-58248-8

Figure Lengend Snippet: Silencing BMI1 enhanced the regulatory effect of CBX7 on the Nrf2/ARE signaling pathway in HGF-induced hepatocytes. ( A , B ) The RT-qPCR was used to assay the expression of CBX7 and BMI1 in NCTC 1469 cells induced by different concentrations of hepatocyte growth factor (HGF, 5, 10, and 20 ng/ml). ( C – G ) Nuclear Nrf2, Nrf2, HO-1, and NQO-1 expression in NCTC 1469 and BNL CL.2 cells was tested by Western blot. β-actin and lamin B was loading control. Values are means ± SD, *P < 0.05, **P < 0.01 vs HGF-0 or pcDNA-NC; # P < 0.05, ## P < 0.01 vs pcDNA-CBX7 + si-NC.

Article Snippet: Mouse normal hepatocytes NCTC 1469 (CL-0407), were obtained from ProCell (Wuhan, China).

Techniques: Quantitative RT-PCR, Expressing, Western Blot

Journal: Open Life Sciences

Article Title: SPOP regulates the expression profiles and alternative splicing events in human hepatocytes

doi: 10.1515/biol-2022-0755

Figure Lengend Snippet: Overexpression of SPOP could facilitate the proliferation of human normal hepatocytes (WRL68). (a) Transfection efficiency of SPOP in WRL68 cells was measured by qRT-PCR. (b) Proliferative abilities of WRL68 cells in the SPOP group and the vec group were detected by CCK8 assay. (c) Colony formation assay detected the proliferations of WRL68 cells in the SPOP group and the vec group. * p < 0.05, ** p < 0.01.

Article Snippet: Human normal hepatocytes WRL68 (ATCC, Manassas, USA) were cultured in Dulbecco’s Modified Eagle Medium (cat. no. PM150210, Procell, Wuhan, China) with 10% GibcoTM fetal bovine serum (cat. no. 10099141C, Invitrogen, AUS) and 1% penicillin–streptomycin (10,000 U/mL) (cat. no. 15140122, Invitrogen, USA), at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Over Expression, Transfection, Quantitative RT-PCR, CCK-8 Assay, Colony Assay

Journal: Open Life Sciences

Article Title: SPOP regulates the expression profiles and alternative splicing events in human hepatocytes

doi: 10.1515/biol-2022-0755

Figure Lengend Snippet: Overexpression of SPOP could change the gene expression profiles of WRL68. (a) Boxplot of sample gene expression distribution. (b) Correlation between SPOP overexpression (SPOP rep1/2/3) and control (vec rep1/2/3) samples. (c) DEGs between the SPOP group and the vec group. (d) Correlation of log2 fold-change between RNA-Seq and RT-qPCR for significantly DEGs.

Article Snippet: Human normal hepatocytes WRL68 (ATCC, Manassas, USA) were cultured in Dulbecco’s Modified Eagle Medium (cat. no. PM150210, Procell, Wuhan, China) with 10% GibcoTM fetal bovine serum (cat. no. 10099141C, Invitrogen, AUS) and 1% penicillin–streptomycin (10,000 U/mL) (cat. no. 15140122, Invitrogen, USA), at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Over Expression, Gene Expression, Control, RNA Sequencing, Quantitative RT-PCR

Journal: Open Life Sciences

Article Title: SPOP regulates the expression profiles and alternative splicing events in human hepatocytes

doi: 10.1515/biol-2022-0755

Figure Lengend Snippet: Potential biological functions of SPOP-related genes in WRL68 cells. (a) Top 30 most enriched GO terms of the upregulated DEGs. (b) Top 30 most enriched GO terms of the downregulated DEGs. (c) Top 20 most enriched KEGG pathways of all DEGs. (d) Top 20 most enriched DisGeNET lists of all DEGs.

Article Snippet: Human normal hepatocytes WRL68 (ATCC, Manassas, USA) were cultured in Dulbecco’s Modified Eagle Medium (cat. no. PM150210, Procell, Wuhan, China) with 10% GibcoTM fetal bovine serum (cat. no. 10099141C, Invitrogen, AUS) and 1% penicillin–streptomycin (10,000 U/mL) (cat. no. 15140122, Invitrogen, USA), at 37°C in a 5% CO 2 humidified atmosphere.

Techniques:

Journal: Open Life Sciences

Article Title: SPOP regulates the expression profiles and alternative splicing events in human hepatocytes

doi: 10.1515/biol-2022-0755

Figure Lengend Snippet: Enrichment biological function of SPOP RASEs in WRL68 cells. (a) Number of significant RASEs. (b) GO functional enrichment of different RASEs. (c) KEGG pathway enrichment analysis results: A3SS events, 11 A5SS events, 20 MXE events, 11 RI events, and 100 SE events.

Article Snippet: Human normal hepatocytes WRL68 (ATCC, Manassas, USA) were cultured in Dulbecco’s Modified Eagle Medium (cat. no. PM150210, Procell, Wuhan, China) with 10% GibcoTM fetal bovine serum (cat. no. 10099141C, Invitrogen, AUS) and 1% penicillin–streptomycin (10,000 U/mL) (cat. no. 15140122, Invitrogen, USA), at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Functional Assay

Journal: Open Life Sciences

Article Title: SPOP regulates the expression profiles and alternative splicing events in human hepatocytes

doi: 10.1515/biol-2022-0755

Figure Lengend Snippet: HCC pathway modified by SPOP overexpression in WRL68 (the figure was drawn according to the information from KEGG PATHWAY database. The molecules in red were upregulated with SPOP overexpression and the molecules in green were downregulated with SPOP overexpression).

Article Snippet: Human normal hepatocytes WRL68 (ATCC, Manassas, USA) were cultured in Dulbecco’s Modified Eagle Medium (cat. no. PM150210, Procell, Wuhan, China) with 10% GibcoTM fetal bovine serum (cat. no. 10099141C, Invitrogen, AUS) and 1% penicillin–streptomycin (10,000 U/mL) (cat. no. 15140122, Invitrogen, USA), at 37°C in a 5% CO 2 humidified atmosphere.

Techniques: Modification, Over Expression