Journal: Pharmaceuticals

Article Title: Acquired Drug Resistance Enhances Imidazoquinoline Efflux by P-Glycoprotein

doi: 10.3390/ph14121292

Figure Lengend Snippet: ( A ) MDR cancer cells were derived from parent cancer cell lines by incrementally increasing the Dox concentration in growth media. ( I ) TRAMP-C2 (TC2) prostate parent cancer cell line. ( II ) B16-F10 (B16) melanoma parent cancer cell line. ( III ) 4T1-Luc2 (4T1) breast parent cancer cell line. ( IV ) TC2-MDR cancer cell line. ( V ) B16-MDR cancer cell line. ( VI ) 4T1-MDR cancer cell line. Scale: 200 µm ( B ) Western blot shows the increased expression of P-gp in MDR cancer cells compared to parent cancer cell lines. ( C ) TC2-Parent, TC2-MDR, B16-Parent, and B16-MDR stained with Anti-P-gp-Alexa Fluor 488 antibody conjugate reveals the increased expression of P-gp in MDR-derived cancer cell lines, as well as membrane localization of P-gp.

Article Snippet: As per manufacturer instructions, the TRAMP C2 cell line (ATCC, Manassas, VA, USA, CRL-2731, mouse transgenic adenocarcinoma) was grown in complete culture media composed of DMEM with 4.5 g L −1 glucose, 2 mM L-glutamine, 100 U mL −1 PenStrep, and supplemented with 1 μg mL −1 Insulin (Sigma-Aldrich, St. Louis, MO, USA, I0516), 2 nM (+)-Dehydroisoandrosterone (VWR, Radnor, PA, USA, 200008-124), 5% HI-FBS, and 5% Nu-Serum IV (Corning, Corning, NY, USA, 355504).

Techniques: Derivative Assay, Concentration Assay, Western Blot, Expressing, Staining, Membrane

Journal: Frontiers in Cell and Developmental Biology

Article Title: Oncostatin M Maintains Naïve Pluripotency of mESCs by Tetraploid Embryo Complementation (TEC) Assay

doi: 10.3389/fcell.2021.675411

Figure Lengend Snippet: OSM maintains telomere elongation, telomerase activity and heterogeneity in 2-cell gene expression. (A) Representative images displaying telomere Q-FISH of mESCs cultured in -LIF, LIF, and OSM medium. Blue, chromosomes stained with DAPI; Red dots, telomeres. Bottom panel, the ratio of normal karyotype in the three culture conditions. (B) Telomere Q-FISH assay showing telomere length distribution by telomere fluorescence unit (TFU). Data shown as Mean ± SD. (C) Expression of telomerase-related genes: Tert and Terc in mESCs cultured in -LIF, LIF and OSM medium. ** p < 0.01; *** p < 0.001; ns, no significant difference. (D) Telomerase activity measured by TRAP assay. Lysis buffer served as negative control. (E) Telomerase activity measured by ELISA assay. A49 mESCs ( Terc –/– G4 ESCs) and N33 mESCs (wild type ESCs) served as negative control and positive control, respectively. * p < 0.05; ** p < 0.01. (F) Relative expression of 2-cell genes including Tcstv1 , Tcstv3 , and Zscan4 in ESCs cultured in -LIF, LIF, and OSM medium. *** p < 0.001; ns, no significant difference. (G) Immunofluorescence staining of Zscan4 (green) and Oct4 (red) in ESCs cultured in -LIF, LIF, and OSM medium. Blue, DAPI. Scale bar, 20 μm. (H) Percentage of Zscan4 + cells (number of mESCs counted in immunofluorescence staining assay). (I) Flow cytometry diagram indicating percentage of Zscan4 + cells in mESCs cultured in -LIF, LIF, and OSM medium. Samples lacking the antibody served as a negative control.

Article Snippet: Telomerase level was determined by ELISA method according to the manufacturer’s instruction using Mouse Telomerase (TE) ELISA kit (CSB-E08022m, CUSABIO).

Techniques: Activity Assay, Gene Expression, Cell Culture, Staining, Fluorescence, Expressing, TRAP Assay, Lysis, Negative Control, Enzyme-linked Immunosorbent Assay, Positive Control, Immunofluorescence, Flow Cytometry