Fig. 7

Interactions between CCRR and CIP85 and between CIP85 and CX43. a RNA immunoprecipitation (RIP) showing the physical interaction between CCRR and CIP85 proteins in cultured neonatal mouse ventricular myocytes (NMVMs). Most left panel: the electrophoresis gel image showing the anticipated fragment representing CCRR that was pulled down by CIP85 antibody with IgG antibody as a negative control. Middle left panel: the quantified results of the electrophoretic bands. ^**P < 0.01 anti-CIP85 vs. anti-IgG; n = 4 batches of cells for each group. Middle right panel: an example of immunoblotting bands showing the pulldown of CIP85 protein by CCRR or other RNAs as labeled on the left. ZFAS1: an lncRNA used as a negative control; AS: antisense to CCRR used as another negative control; AR: androgen receptor 3′ UTR: a commercially obtained positive control; Input: purified CIP85 as a positive control; NC: negative control probe; HuR: Hu-antigen R. Right panel: mean relative band densities representing CIP85 protein with varying treatments. ^***P < 0.001 AS, ZFAS1, NC vs. CCRR; n = 4. b Protein co-immunoprecipitation (Co-IP) confirming the physical interaction between CIP85 and Cx43 in cultured NMVMs. Upper panel shows the presence of CIP85 in an anti-Cx43 pulldown sample, and lower panel shows the presence of Cx43 in an anti-CIP85 pulldown sample. c RIP results showing the physical interaction between FD and CIP85 proteins in heathy mice. Right panel: the quantified results of the electrophoretic bands. ^**P < 0.01 anti-CIP85 vs. anti-IgG; n = 3 mice for each group. d Immunoblotting image showing the pulldown of CIP85 protein by FD. Sham: non-treated sample; Lv-FD: lentiviral vector carrying FD. Lv-FD-NC: lentiviral vector carrying negative sequence. Lower panel: mean relative band densities representing CIP85 protein with varying treatments. ^**P < 0.01 Lv-FD vs. Lv-FD-NC; n = 4. (Mean ± SEM; ANOVA followed by Dunnett’s test for multiple group comparisons, and Student t test for two group comparisons)