Figure 4
From: Interaction of Cx43 with Hsc70 regulates G1/S transition through CDK inhibitor p27
Cx43 competes with p27 for binding to Hsc70.
(a) A schematic representation of Hsc70 and its deletion mutants is shown. (b,c) Identification of Cx43 (b) or p27 (c) binding domain in Hsc70. HuH-7 cells were transfected with expression vectors encoding Cx43 or HA-p27 and various forms of Hsc70 used in co-immunoprecipitation assays. The immunocomplexes were analyzed on 12% SDS-PAGE followed by western blotting (WB) using anti-T7-tag and anti-Cx43 or anti-HA-tag antibodies. Cropped blots are shown. (d) Cx43 and p27 competitively bound to Hsc70. HuH-7 cells were transfected with equivalent plasmids of T7-Hsc70 and HA-p27 and with increasing amounts of Cx43 as indicated in the figure. Lysates were immunoprecipitated by anti-T7 antibody. The immunocomplexes were analyzed on 12% SDS-PAGE followed by WB using anti-T7-tag, anti-Cx43 and anti-HA-tag antibodies. Cropped blots are shown and full-length blots are presented in Supplementary Fig. S2d.
