Fig 1: Effect of PRAF2 on the Wnt/ß-catenin signaling pathway. (A) Western blotting of PRAF2, ß-catenin and TCF4 proteins in MCF-7 cells. (B) Semi-quantitative analysis of protein expression of PRAF2, ß-catenin and TCF4 in MCF-7 cells. (C) mRNA expression of PRAF2, ß-catenin and TCF4 in MCF-7 cells detected by reverse transcription-quantitative PCR. *P<0.05. PRAF2, prenylated rab acceptor 1 domain family member 2; si, small interfering; NC, negative control; TCF4, T-cell factor 4.
Fig 2: PRAF2 expression in breast cancer and normal adjacent tissues. Expression of PRAF2 in (A) BRCA samples and normal breast tissues, (B) BRCA samples at different cancer stages and (C) BRCA samples with different conditions of nodal metastasis, based on data from TCGA database. (D) Quantitative analysis of the mRNA expression of PRAF2 in T and N tissues as detected by reverse transcription-quantitative PCR. (E) Representative western blots of the protein expression of PRAF2 in T and N tissues, normalized to ß-actin. (F) Semi-quantitative analysis of protein expression of PRAF2 in T and N tissues by western blotting (n=37). ***P<0.001 compared with normal. BRCA, breast cancer; N, normal; T, tumor; PRAF2, prenylated rab acceptor 1 domain family member 2; TCGA, The Cancer Genome Atlas.
Fig 3: Western blot analysis detection of the siRNA knockdown efficiency of PRAF2. (A) Expression of PRAF2 mRNA in breast cancer cell lines MCF-7 and MDA-MB-231 and normal mammary epithelial cell line MCF-10A detected by RT-qPCR. (B) Expression of PRAF2 protein in MCF-7, MDA-MB-231 and MCF-10A cell lines detected by western blotting and (C) semi-quantitative analysis of the protein expression of PRAF2. (D) PRAF2 expression in MCF-7 cells transfected with siRNA-PRAF2 detected by RT-qPCR. (E) PRAF2 expression in MCF-7 cells transfected with siRNA-PRAF2 detected by western blotting and (F) semi-quantitative analysis of the protein expression of PRAF2. ***P<0.001. PRAF2, prenylated rab acceptor 1 domain family member 2; RT-qPCR, reverse transcription-quantitative PCR; si, small interfering; NC, negative control.
Fig 4: Effect of PRAF2 on the proliferation of MCF-7 cells as assessed by Cell Counting Kit-8 and colony formation assays. (A) Representative images from colony formation assay in MCF-7 cells. (B) A statistically significant difference in the colony number was found between the siRNA-NC and siRNA-PRAF2 groups. (C) A statistically significant difference in the proliferation rate was found between siRNA-NC and siRNA-PRAF2 MCF-7 cells. *P<0.05; ***P<0.001. PRAF2, prenylated rab acceptor 1 domain family member 2; si, small interfering; NC, negative control.
Fig 5: Effect of PRAF2 on the migration of MCF-7 cells as detected by scratch assay. (A) Representative images of wound healing assay of MCF-7 cells at 0 and 24 h after transfection. Scale bars, 500 µm. (B) A statistically significant difference in cell migration was found between siRNA-NC and siRNA-PRAF2 MCF-7 cells. ***P<0.001. PRAF2, prenylated rab acceptor 1 domain family member 2; si, small interfering; NC, negative control.
Supplier Page from Abcam for Anti-JM4/PRAF2 antibody