Fig 1: RNF6 promotes carboplatin resistance through JAK2/STAT3 signaling pathway. (A) Carboplatin-resistant RB cells Y-79/CR were first treated with carboplatin in the presence or absence of various signaling pathway inhibitors for 24 h, and then cell viability was assessed by MTT assay. Cells without signaling pathway inhibitor treatment were used as control and were considered 100% viable. Data shown are mean ±SD of three independent experiments. **, p < 0.01. (B) Carboplatin-resistant RB cells Y-79/CR were first transfected with control siRNA, STAT3 siRNA or NF-κB siRNA, and then treated with carboplatin for 24 h. After treatment, cell viability was assessed by MTT assay. Cells without siRNA transfection were used as control and were considered 100% viable. Data shown are mean ±SD of three independent experiments. **, p < 0.01. (C) Carboplatin-resistant RB cells Y-79/CR were first transfected with control siRNA, STAT3 siRNA or NF-κB siRNA, and then treated with carboplatin for 24 h. After treatment, the expression of RNF6, p65, STAT3 and p-STAT3 was determined by western blot. One representative result is shown. (D) Densitometry analysis was performed for (C). *, p < 0.05; **, p < 0.01.
Fig 2: RNF6 promotes CC cell growth. (A) The protein level of RNF6 was measured by western blotting. *P < 0.05 (B) The transfection efficiency was measured by qRT‐PCR. *P < 0.05 (a) RNF6 siRNA‐1, RNF6 siRNA‐2, or si‐NC was transfected into HeLa cells. (b) RNF6 overexpression plasmid or empty plasmid was transfected into HeLa cells. (C) MTT assays was used to assess cell viability at certain points in time. (Mean ± SD; n = 4; Student’s t‐test and one‐way ANOVA). *P < 0.05.
Fig 3: Expression of RNF6 is significantly upregulated in BC tissue. (A) RT-qPCR analysis of RNF6 mRNA expression in 146 BC tumor and paired non-tumor tissue samples. **P<0.01. (B) Western blotting and (C) quantification of RNF6 protein levels in BC and paired non-tumor tissue. Tubulin was used as a loading control. (D) Representative images of RNF6 staining. Magnification, x400, Scale bar, 100 µm. (E) Western blotting and (F) RT-qPCR were performed to detect RNF6 protein and mRNA levels in four BC cell lines and MCF10A cells. *P<0.05, **P<0.01 and ***P<0.001 vs. MCF10A. RNF6, ring finger protein 6; BC, breast cancer; RT-q, reverse transcription-quantitative.
Fig 4: RNF6 overexpression promotes carboplatin resistance in retinoblastoma cells. RB cells (A) Y-79 and (B) SO-Rb50 were first transfected with control plasmid pcDNA3.1 or RNF6-coding plasmid pcDNA-RN6, and then cells were treated with carboplatin for 24 h. After treatment, cell viability was assessed by MTT assay. Cells with neither plasmid transfection nor carboplatin treatment were used as control and were considered 100% viable. Data shown are mean ±SD of three independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.
Fig 5: RNF6 inhibits apoptosis of CC cells. (A) Cell apoptosis was promoted after knockdown of RNF6 in HeLa cells. Quadrants: upper left (necrotic cells); lower left (live cells); lower right (early apoptotic cells); and upper right (late apoptotic cells). (B) Cell apoptosis was inhibited after RNF6 overexpression. (Mean ± SD; n = 4; Student’s t‐test; and one‐way ANOVA). *P < 0.05.
Supplier Page from Abcam for Anti-RNF6 antibody