Figure 3
From: Interaction of Cx43 with Hsc70 regulates G1/S transition through CDK inhibitor p27
Association among Cx43, Hsc70 and p27.
(a) Hsc70 interacted with Cx43. Lysates prepared from HuH-7 cells transfected with T7-Hsc70 and wild-type Cx43 were immunoprecipitated by anti-Cx43 antibody or control IgG. The immunocomplexes were analyzed on 12% SDS-PAGE followed by western blotting (WB) using anti-T7 antibody. (b) Hsc70 interacted with p27. Lysates prepared from HuH-7 cells transfected with T7-Hsc70 and HA-p27 were immunoprecipitated by anti-T7-tag antibody or control IgG. The immunocomplexes were analyzed on 12% SDS-PAGE followed by WB using anti-HA-tag antibody. (c) Absence of interaction between Cx43 and p27. Lysates prepared from HuH-7 cells transfected with both wild-type Cx43 and HA-p27 expression plasmids were immunoprecipitated by anti-Cx43 antibody or control IgG. The immunocomplexes were analyzed on 12% SDS-PAGE followed by WB using anti-HA antibody. (a–c) Cropped blots were shown and full-length blots are presented in Supplementary Fig. S2a, S2b and S2c, respectively. (d) Partial subcellular co-localization of p27 and Hsc70. HuH-7 cells were transfected with HA-p27 and T7-Hsc70. Cells were fixed and stained with anti-HA-tag and anti-T7-tag antibodies. (e) Partial subcellular co-localization of Cx43 and Hsc70. HuH-7 cells were transfected with Cx43 and T7-Hsc70. Cells were fixed and stained with anti-Cx43 and anti-T7-tag antibodies.
