rabbit polyclonal anti survivin af886  (R&D Systems)

Journal: International Journal of Medical Sciences

Article Title: CD44+/CD105+ Human Amniotic Fluid Mesenchymal Stem Cells Survive and Proliferate in the Ovary Long-Term in a Mouse Model of Chemotherapy-Induced Premature Ovarian Failure

doi: 10.7150/ijms.4841

Figure Lengend Snippet: Analysis of the proliferation and survival of CD44+/CD105+ HuAFCs in vitro . (A) MTT assays indicated that the viability of CD44-/CD105- HuAFCs significantly reduced at both 2 d and 5 d, compared to CD44+/CD105+ HuAFCs. ** P < 0.01 vs. CD44-/CD105- HuAFCs; * P < 0.05 vs. CD44-/CD105- HuAFCs; # P > 0.05 vs. CD44-/CD105- HuAFCs; n = 3. (B) qRT-PCR analysis of Ki67 and survivin mRNA expression in CD44+/CD105+ and CD44-/CD105- HuAFCs; * P < 0.05 vs. CD44-/CD105- HuAFCs; # P > 0.05 vs. CD44-/CD105- HuAFCs; n = 3. (C) Western blotting analysis of Ki67 and survivin protein expression in CD44+/CD105+ and CD44-/CD105- HuAFCs; ** P < 0.01 vs. CD44+/CD105+ HuAFCs; * P < 0.05 vs. CD44+/CD105+ HuAFCs; # P > 0.05 vs. CD44+/CD105+ HuAFCs; n = 3. (D) Flow cytometric cell cycle analysis of CD44+/CD105+ and CD44-/CD105- HuAFCs. The majority of CD44-/CD105- HuAFCs were arrested in the G2/M phase with a reduced percentage of S phase cells; ** P < 0.01 vs. CD44+/CD105+ HuAFCs; * P < 0.05 vs. CD44+/CD105+ HuAFCs; # P > 0.05 vs. CD44+/CD105+ HuAFCs; n = 3. (E) FCM analysis of human mesenchymal stem cell marker expression in CD44+/CD105+ and CD44-/CD105- HuAFCs in vitro. Expression of the "stemness" markers was higher in CD44+/CD105+ HuAFCs than CD44-/CD105- HuAFCs.

Article Snippet: After blocking with 5% (w/v) non-fat dried milk in TBST (Tris-buffered saline containing Tween-20) (25 mM Tris/HCl, pH 8.0, 125 mM NaCl and 0.05% Tween-20), the PVDF membranes were washed 4 times (15 min each) with TBST at room temperature and incubated with primary antibody (rabbit anti-human Ki67 polyclonal antibody (1:1000; Chemicon, Temecula, CA, USA) and rabbit anti-human Survivin polyclonal antibody (1:1000; Chemicon, Temecula, CA, USA)).

Techniques: In Vitro, Quantitative RT-PCR, Expressing, Western Blot, Cell Cycle Assay, Marker

Journal: International Journal of Molecular Sciences

Article Title: Arachidin-1, a Prenylated Stilbenoid from Peanut, Induces Apoptosis in Triple-Negative Breast Cancer Cells

doi: 10.3390/ijms23031139

Figure Lengend Snippet: Western blot analysis and densitometry measurements of treated TNBC cell lines with arachidin-1 (A-1, 0.1, and 1 µM). Western blot images of ( A ) PARP and cleaved PARP, GAPDH loading control. ( B ) Full-length caspase-8, GAPDH loading control; ( C ) cleaved caspase-9, vinculin loading control; and ( D ) survivin, GAPDH loading control. Lysates (60 µg of total protein) of MDA-MB-231 and MDA-MB-436 cells treated with A-1 were analyzed by western blotting. The relative densitometry of ( E ) full-length PARP, ( F ) cleaved PARP, ( G ) caspase-8, ( H ) cleaved caspase-9, and ( I ) survivin protein levels were analyzed by comparing to the loading control. Cells with 0.01% DMSO were used as controls. Data represents mean ± SD from three or more independent experiments. * p < 0.05, ** p < 0.001, **** p < 0.0001 versus control in MDA-MB-231, # p < 0.05, ## p < 0.001, #### p < 0.00001 versus control in MDA-MB-436, and ns = not significant from control.

Article Snippet: The primary antibodies and dilutions included rabbit anti-caspase-8 (D35G2) mAb (1:500; Cell Signaling Technology; Danvers, MA, USA), rabbit anti-PARP mAb (1:500; Cell Signaling Technology; Danvers, MA, USA), rabbit anti-human survivin polyclonal antibody (0.5 μg/mL; R&D Systems; Minneapolis, MN, USA), mouse anti-human caspase-9 mAb (1 μg/mL; R&D Systems; Minneapolis, MN, USA), mouse anti-vinculin antibody (2 μg/mL; R&D Systems; Minneapolis, MN, USA), and mouse anti-human GAPDH antibody (0.05 μg/mL; R&D Systems; Minneapolis, MN, USA).

Techniques: Western Blot, Control