Journal: Oncology Reports

Article Title: Filamin B extensively regulates transcription and alternative splicing, and is associated with apoptosis in HeLa cells

doi: 10.3892/or.2020.7532

Figure Lengend Snippet: Knockdown of FLNB promotes proliferation and inhibits apoptosis of HeLa cells. (A) Relative mRNA expression of FLNB in HeLa cells after it was knocked down using FLNB -specific shRNA was determined by reverse transcription-qPCR. shFLNB_1 and shFLNB_2 indicate two biological repeats of HeLa cells transfected with FLNB-specific shRNA. Ctrl_1 and Ctrl_2 indicate two biological repeats of HeLa cells transfected with empty vector as controls. (B) Cell proliferation of shFLNB was measured by an MTT assay in HeLa cells. Cell apoptosis of cells transfected with shFLNB was measured by (C) flow cytometry and (D) subsequent analysis by a 7-ADD and Annexin V assay. *P<0.05 and **P<0.01 vs. respective Ctrl. FLNB , filamin B; sh, short hairpin; qPCR, quantitative PCR; Ctrl, control; 7-AAD, 7-amino actinomycin D; OD, optical density; PE, phycoerythrin.

Article Snippet: Subsequently, 5% bovine serum albumin (Gibco; Thermo Fisher Scientific, Inc.) was selected to block the membranes for 1 h at room temperature, which were then incubated with primary antibodies against FLNB (ABclonal Biotechnology Co., Ltd.; 1:500, cat. no. A2481), NLR family apoptosis inhibitory protein (NAIP; BIOSS; 1:1,000; cat. no. bs-5804R), mitogen-activated protein kinase kinase 7 (MAP2K7; ABclonal Biotechnology Co., Ltd.; 1:1,000; cat. no. A2186) and GAPDH (ABclonal Biotechnology Co., Ltd.; 1:50,000; cat. no. AC036) at 4°C overnight with gentle rocking.

Techniques: Expressing, shRNA, Transfection, Plasmid Preparation, MTT Assay, Flow Cytometry, Annexin V Assay, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Filamin B extensively regulates transcription and alternative splicing, and is associated with apoptosis in HeLa cells

doi: 10.3892/or.2020.7532

Figure Lengend Snippet: Validation of FLNB -regulated genes (DEGs). (A) Relative expression level (FPKM, up) and RT-qPCR measurement (down) of cartilage development-related DEGs. (B) Related expression level (FPKM, up) and RT-qPCR measurement (down) of apoptotic-related DEGs. (C) Western blot analysis of two apoptotic-related proteins in shFLNB and Ctrl HeLa cells. FLNB , filamin B; DEGs, differentially expressed genes; FPKM, fragments per kilobase of transcript per million fragments mapped; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin; Ctrl, control; ATP7A, ATPase copper transporting α; BMP7, bone morphogenetic protein 7; COL2A1, collagen type II α 1 chain; MMP13, matrix metallopeptidase 13; IL23A, interleukin 23 subunit α; MALAT1, metastasis associated lung adenocarcinoma transcript 1; NAIP, NLR family apoptosis inhibitory protein; SLC25A36, solute carrier family 25 member 36; MAP2K7, mitogen-activated protein kinase kinase 7.

Article Snippet: Subsequently, 5% bovine serum albumin (Gibco; Thermo Fisher Scientific, Inc.) was selected to block the membranes for 1 h at room temperature, which were then incubated with primary antibodies against FLNB (ABclonal Biotechnology Co., Ltd.; 1:500, cat. no. A2481), NLR family apoptosis inhibitory protein (NAIP; BIOSS; 1:1,000; cat. no. bs-5804R), mitogen-activated protein kinase kinase 7 (MAP2K7; ABclonal Biotechnology Co., Ltd.; 1:1,000; cat. no. A2186) and GAPDH (ABclonal Biotechnology Co., Ltd.; 1:50,000; cat. no. AC036) at 4°C overnight with gentle rocking.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Real-time Polymerase Chain Reaction