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Fig. 1. RPB3 subcellular localization. (A) Endogenous RPB3 in cycling (GM) and differentiated (DM) myogenic C2C7 cells. (B) Endogenous RPB3 in HeLa cells. (C) Transfected Myc-RPB3 in cycling (GM) and differentiated (DM) myogenic C2C7 cells. (D) Endogenous RPB11 in cycling (GM) and differentiated (DM) myogenic C2C7 cells. Indirect immunofluorescence (A-D) was performed with the indicated antibodies. Hoechst-stained nuclei relative to the same fields are shown. (E) Western-blot analysis of subcellular cytoplasmic and nuclear fractions derived from GM and DM myogenic C2C7 cells. The blot was incubated with anti-RPB3 antibody to determine nuclear/cytoplasmic localization. To verify fraction quality, the same extracts were blotted and incubated with anti-Sp1 antibody.