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: Human HeLa cells (CCTCC@GDC0009) were obtained from The China Center for Type Culture Collection.

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

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: GO and KEGG analysis of differentially expressed genes between short hairpin FLNB-transfected and control HeLa cells. Top 10 most enriched GO terms (biological process) of (A) upregulated and (B) downregulated genes upon FLNB knockdown. Rectangles around GO terms indicate notable cancer-related and cartilage development terms. Top 10 most enriched KEGG pathways of (C) upregulated and (D) downregulated genes upon FLNB knockdown. GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; FLNB , filamin B.

Article Snippet: Human HeLa cells (CCTCC@GDC0009) were obtained from The China Center for Type Culture Collection.

Techniques: Transfection, Control, Knockdown

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: Human HeLa cells (CCTCC@GDC0009) were obtained from The China Center for Type Culture Collection.

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Control

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 -affected ASEs. Genome visualization (left panel) shows FLNB -regulated ASEs in shFLNB and control. (A) Validation of an ASE of VDR in HeLa cells. (B) Validation of an ASE of PACS2 in HeLa cells. (C) Validation of an ASE of MAP2K7 in HeLa cells. (D) Validation of an ASE of MALAT1 in HeLa cells. The number of junction reads were marked on the line representing splice junction composing ASE. The structures of ASEs are depicted in the top-right panel. The altered ratio of ASEs in RNA-sequencing and in reverse transcription-quantitative PCR were calculated and plotted (right panel, bottom). FLNB , filamin B; ASEs, alternative splicing events; sh/SH, short hairpin; NC, negative control; VDR, vitamin D receptor; PACS2, phosphofurin acidic cluster sorting protein 2; MAP2K7, mitogen-activated protein kinase kinase 7; MALAT1, metastasis associated lung adenocarcinoma transcript 1. *P<0.05 and ***P<0.001 vs. respectively NC.

Article Snippet: Human HeLa cells (CCTCC@GDC0009) were obtained from The China Center for Type Culture Collection.

Techniques: Biomarker Discovery, Control, RNA Sequencing, Reverse Transcription, Real-time Polymerase Chain Reaction, Alternative Splicing, Negative Control

Journal: Scientific Reports

Article Title: Mechanisms and immunogenicity of nsPEF-induced cell death in B16F10 melanoma tumors

doi: 10.1038/s41598-018-36527-5

Figure Lengend Snippet: Histological analysis of nsPEF treated tumors. 30–50 mm 3 B16F10 tumors were treated with 750, 200-ns pulses (25 kV/cm, 2 Hz) or left untreated (sham control). Panel A shows H&E pictures for one sham and one nsPEF-treated tumor collected at 4 h post treatment. In ( B ), both anti-cleaved caspase 3 (green) and -Ki-67 (red) immunofluorescence were performed to assess apoptosis and cell proliferation, respectively. Panel B shows representative images from three sham (top) and three nsPEF (bottom) -treated tumors. Panel C shows a positive control for the anti-cleaved Caspase 3 staining, namely HeLa cells treated with 1 μm staurosporin for 5 h. Scale bar: 1000 μm or 100 μm (inset) ( A ); 100 μm ( B , C ).

Article Snippet: Mouse melanoma B16F10 (ATCC, Manassas, Virginia) and human cervix carcinoma HeLa cell lines were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Corning®, Corning, NY) while the human lymphoma U-937 cell line was cultured in RPMI 1640 (Corning®).

Techniques: Immunofluorescence, Positive Control, Staining

Journal: Scientific Reports

Article Title: On the mechanism of miR-29b enhancement of etoposide toxicity in vitro

doi: 10.1038/s41598-024-70856-y

Figure Lengend Snippet: Effect of siRNA against Mcl-1 on etoposide cytotoxicity in different cell lines. (Panel a ): The cells were transfected (6 h) with siRNA against Mcl-1 or negative control. Cells were then treated with 60 μM etoposide 24 h post transfection and incubation continued until experiment was ended. The proliferation was calculated from doubling time value as explained in Materials and methods section. Each bar represents mean ± s.d. of at least three independent experiments (N HeLa = 4, N Hep G2 = 3, N Caco-2 = 3), in each experiment triplicate measurements were performed. **p < 0.01 versus corresponding negative control. (Panel b ): Evaluation of siRNA efficiency by western blot analysis in Hela cells. The cells were transfected (6 h) with anti-Mcl-1 siRNA or negative control. Shown are representative western blots of cells cultured for 24 h after transfection. (Panel c ): Evaluation of siRNA efficiency by western blot analysis in Hep G2 cells. Treatment was the same as for panel b. Panel d: Evaluation of siRNA efficiency by western blot analysis in Caco-2 cells. Treatment was the same as for (panel b ).

Article Snippet: HeLa cells were obtained from The European Collection of Authenticated Cell Cultures (ECACC, No. 93021013 Human Negroid cervix epitheloid carcinoma cell line) via Sigma-Aldrich as provider.

Techniques: Transfection, Negative Control, Incubation, Western Blot, Cell Culture