Figure 4
From: Maspin overexpression modulates tumor cell apoptosis through the regulation of Bcl-2 family proteins
Changed levels of Bcl-2 family proteins in TC, T16, and T18 tumor cells. A. Analysis of anti-apoptotic Bcl-2 proteins in tumor cells. Cells were cultured for 48 hrs in serum starvation condition before they were lysed in cell lysis buffer. Cytosolic proteins (1 mg) were immunoprecipitated with Bcl-2 and Bcl-XL monoclonal antibodies overnight at 4°C. The level of Bcl-2 and Bcl-XL proteins was detected using a HRP-conjugated anti-mouse secondary antibody. IgG serves as an internal control. Right panel, relative level of Bcl-2 and Bcl-XL proteins in TC, T16, and T18 cells. The protein bands were quantitated using a densitometer and were normalized to IgG signals. B. Analysis of pro-apoptotic Bax protein in TM40D cells. Cells were induced to apoptosis with serum starvation. Cell extracts were run on SDS PAGE gel and immunobloted with anti-Bax and anti-beta-actin antibodies. Beta-actin serves as a loading control. Right panel, relative level of Bax protein in TC, T16, and T18 cells. The protein bands were quantitated using a densitometer and were normalized to beta-actin signal.