Figure 4.

RP11-147L13.8interacts with c-Jun and suppresses the c-Jun-ser73 phosphorylation via inhibiting the JNK in GBC cells

(A) RP11-147L13.8 pull-down assay analyzed by SDS-PAGE. (B) The RP11-147L13.8-c-Jun interaction was verified by RNA immunoprecipitation (RIP) assays, wherein RP11-147L13.8 was significantly enriched in c-Jun antibody, but not IgG control. (C) The western blot results, which confirmed that c-Jun protein was specifically associated with sense, but not anti-sense, RP11-147L13.8 in both GBC-SD and NOZ cell lines are shown. (D and E) The overexpression of RP11-147l13.8 could suppress the ser73 phosphorylation of the c-Jun protein, although the knockdown of RP11-147l13.8 could promote the ser73 phosphorylation of the c-Jun protein in both GBC-SD and NOZ cell lines. Data represent mean ± SEM (three biological replicates). (F) The co-immunoprecipitation experiment indicated that the overexpression of RP11-147L13.8 could competitively inhibit the interaction between the JNK and c-Jun protein, which results in the ser73 dephosphorylation of c-Jun protein in both GBC-SD and NOZ cell lines. (G) The co-immunoprecipitation experiment indicated that the knockdown of RP11-147L13.8 could enhance the interaction between the JNK and c-Jun protein, which results in the ser73 phosphorylation of c-Jun protein in both GBC-SD and NOZ cell lines.