FIG 2.

Interaction between RBM24 and Pol is independent of RNA/ε. (A) HepG2 cells were cotransfected with plasmids pHA-Pol and pFlag-RBM24, pFlag-ΔRNP1/2, or the control vector. The cells were immunostained with an anti-HA antibody and an anti-Flag antibody at 48 hpt. Nuclei were stained with Hoechst 33258. Higher-magnification images of the selected areas are also shown. Bars, 10 μm for panels a1 to a4, b1 to b4, c1 to c4, and d1 to d4 and 2 μm for panels a5, b5, c5, and d5. (B) HEK293T cells were cotransfected with HA-Pol, pFlag-RBM24, and pCMV-HE or the empty vector and lysed at 48 hpt. Cell lysates were pretreated with RNase A and DNase I or left untreated and subjected to a co-IP assay. An anti-Flag antibody was used for co-IP. The precipitates were analyzed by Western blotting using anti-Flag and anti-HA antibodies. Nucleic acid was detected by agarose gel electrophoresis. Actin served as a loading control. (C) HEK293T cells were cotransfected with pHA-Pol and pFlag-ΔRNP1/2 or the empty vector. The cells were lysed and subjected to a co-IP assay at 48 hpt. An anti-Flag antibody was used for the co-IP. The precipitates were analyzed by Western blotting using anti-Flag and anti-HA antibodies. Actin served as a loading control.