Skip to main content
. 2008 Jan;10(1):61–68. doi: 10.1593/neo.07885

Figure 5.

Figure 5

Immunoglobulins (IgM) were detected in the cytoplasm of B16F10-Nex2 murine melanoma cells permeabilized with a low dose of gomesin. (A) Chemiluminescent immunoblot analysis showing mAb A4M reactivity with histone 1. NE B16, nuclear extract of B16F10-Nex2 cells; H1 Sigma, commercially purified calf thymus histone from Sigma. Lane 1, 25 µg/ml mAb A4M; Lane 2, 25 µg/ml commercial anti-pan histone antibody; Lane 3, 25 µg/ml an irrelevant mAb. (B) Cells were incubated with a low dose of Gm (2 µM) or with 0.5% saponin and 100 µg/ml mAb A4M for 12 hours. Samples were incubated with FITC-conjugated secondary antibodies and cytoplasmic fluorescence was quantified by FACS. None, untreated cells; Ig-FITC, cells treated with secondary antibodies in the absence of mAbs; FL1-H, fluorescence intensity. (C) Cells were incubated with different concentrations of A4M mAb (25, 50, 80, and 100 µg/ml) in presence (+Gm) or absence (-Gm) of 2 µM Gm for 12 hours. Viable cells were counted in the presence of Trypan blue. None, untreated cells; Gomesin, cells treated with 2 µM Gm. Data are a representative experiment of a triplicate set. Bars represent means and SD.