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

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: Oncotarget

Article Title: Myocardin-related transcription factor A (MRTF-A) activity-dependent cell adhesion is correlated to focal adhesion kinase (FAK) activity

doi: 10.18632/oncotarget.12350

Figure Lengend Snippet: A, E and I. HeLa (cervix carcinoma), HCA7 (colon adenocarcinoma) and SK-UT-1 (uterine leiomyosarcoma) cells were transfected with CA-MRTF-A or empty vector in combination with GFP. Western blotting analysis showed that expression of CA-MRTF-A increased expression levels of MRTF-SRF-dependent FA proteins and the tyrosine phosphorylation levels of FAK and paxillin in HeLa (A), HCA7 (E) and SK-UT-1 (I) cells. B, F and J. Representative images of CA-MRTF-A-transfected and control cells stained with anti-phospho-Tyr397 FAK and anti-phospho-Tyr118 paxillin (red). HeLa (B), HCA7 (F) and SK-UT-1 (J) cells were shown. Transfected cells expressed GFP (green). Scale bar, 20 μm. Insets: high magnification image in grayscale. Scale bar, 10 μm. C, G and K. Migration speed was measured by time-lapse imaging using In Cell Analyzer. CA-MRTF-A (HeLa, n = 5; HCA7, n = 4; SK-UT-1, n = 3). D, H and L. Measurement of the attached cell area during cell migration (HeLa, n = 5; HCA7, n = 4; SK-UT-1, n = 3). Error bars indicate SEM. Paired Student's t-test. *p < 0.05, **p < 0.01.

Article Snippet: HeLa human cervix carcinoma cells and HCA7 human colon adenocarcinoma cells are purchased from Sumitomo Dainippon Pharma and ERACC, respectively.

Techniques: Transfection, Plasmid Preparation, Western Blot, Expressing, Phospho-proteomics, Control, Staining, Migration, Imaging