Figure 5.

DEPDC1B interacts with Rac1 and activates the Rac1/PAK1-LIMK1-cofilin1 signaling pathway. (A) A correlation between DEPDC1B expression and Rac1 expression in PC (data from TCGA). (B) Co-IP experiments using PANC-1 and CFPAC-1 cells confirmed that DEPDC1B and Rac1 can bind with each other. (C) To ensure that the immunoprecipitation procedures worked properly, positive control (GTPγS) and negative control (GDP) experiments were carried out before the formal experiment. (D) GTP-Rac1 (Rac1 activation) levels were measured by GST-PAK1-PBD pull-down assay and were analyzed by Western blotting in the siNC, siDEPDC1B#1, LVCON319 and LV-DEPDC1B groups. The expression level of GTP-Rac1 was quantified relative to the total Rac1 level. (E) The expression of related proteins in the Rac1/PAK1-LIMK1-cofilin1 signaling pathway was assayed by Western blotting in the siNC, siDEPDC1B#1, LVCON319 and LV-DEPDC1B groups. *P <0.05, **P <0.01, and ***P <0.001.

Abbreviations: DEPDC1B, DEP domain-containing protein 1B; PC, pancreatic cancer; TCGA, The Cancer Genome Atlas; Co-IP, Co-immunoprecipitation.