Fig. 2. Leptomycin B (LMB)-sensitive localization of endogenous rpS6 and rpL23a. (A) Specificity of the anti-L23a and anti-S6 antibodies. Total HeLa cell proteins were separated by 8% SDS-PAGE followed by immunoblotting using anti-L23a and anti-S6 antibodies. Note that both antibodies are highly specific and detect a single protein band. (B) HeLa cells were either left untreated or treated with LMB for 18 hours. Then, cells were fixed/permeabilized with either 3.7% paraformaldehyde (PFA)/acetone or with acetone/methanol. The localization of rpL23a or rpS6 was analyzed by indirect immunofluorescence using anti-L23a and anti-S6 antibodies, respectively. Images were taken by confocal fluorescence microscopy.