Journal: Molecular Biology of the Cell
Article Title: Ablation of Nonmuscle Myosin II-B and II-C Reveals a Role for Nonmuscle Myosin II in Cardiac Myocyte Karyokinesis
Figure Lengend Snippet: Increased acetylated tubulin in siRNA NM II-A/II-B–treated HL-1 cells. Immunofluorescence confocal images of B + C + /B + C + HL-1 cells (A–C) and NM II-A/II-B siRNA-treated HL-1 cells (D–F) stained with antibodies to NMHC II-A (A and D, green), p21 (A and D, red), acetylated tubulin (B and E, red), activated caspase-3 (C and F, green), and β-tubulin (C and F, red). NMHC II-A is significantly lower in siRNA-treated cells (D, green) compared with controls (A, green). Although the number of p21-positive cells increases after siRNA treatment, the overall expression level of p21 is not obviously different in siRNA-treated individual cells (D) compared with control cells (A). siRNA-treated HL-1 cells show marked increase in acetylated tubulin (E, red) compared with control cells (B, red). siRNA treatment also increases the number of apoptotic cells detected by activated caspase-3 (F, green) compared with control HL-1 cells (C). DAPI (blue) stains the nuclei.
Article Snippet: The following primary antibodies were used in this study: rabbit polyclonal antibodies NMHC II-A, II-B, and II-C (IF, 1:1000; ; ); connexin43 (IF, 1:100; Cell Signaling Technology, Danvers, MA); and desmin (IF, 1:200; Abcam, Cambridge, MA); and mouse monoclonal antibodies desmin (IF, 1:200; Dako North America, Carpinteria, CA), MF20 (IF, 1:30; Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), E-cadherin (IF, 1:1000; BD Biosciences, San Jose, CA), N-cadherin (IF, 1:200; Zymed Laboratories, South San Francisco, CA), β-catenin (IF, 1:200; Zymed Laboratories), β-actin (IB, 1:5000; Sigma-Aldrich, St. Louis, MO), β-tubulin (IF, 1:1000; Sigma-Aldrich), acetylated α-tubulin (IF, 1:200; Sigma-Aldrich), γ-tubulin (IF, 1:1000; Sigma-Aldrich), and p21 (IB, 1:500; IF, 1:50; BD Biosciences).
Techniques: Immunofluorescence, Staining, Expressing