Fig. 4: ROCK2 regulates dynamin-related protein 1 expression and ROCK2 and Drp1 expression are positively correlated in CCA tissues.

A, B Proteomics sequencing analysis was performed to identify differentially expressed proteins in control and ROCK2-knockdown R-QBC-939 cells. C The expression of Drp1 was detected by Western blot in RBE, R-RBE, QBC-939, and R-QBC-939 cells. D The expression of Drp1 and ROCK2 was detected by western blot in control and Drp1-knockdown R-QBC-939 and R-RBE cells. E–G Cell proliferation was measured using CCK-8 assay, clone formation assay, and EdU assay in control and Drp1-knockdown R-QBC-939 and R-RBE cells after treatment with Pemigatinib (5 μM, 48 h at 37 °C). Data indicated as mean ± SD. ***P < 0.001, **P < 0.01. H The expression of Drp1 and ROCK2 was detected by western blot in control and ROCK2-knockdown R-QBC-939 and R-RBE cells. I–K Representative IHC staining of ROCK2 and Drp1 in pCCA, dCCA and iCCA. (top: ×100 magnification, scale bar, 200 μm; bottom: ×400 magnification, scale bar, 50 μm). L Patients were divided into subsets with low or high Drp1 expression according to the Remmele score. The ROCK2 Remmele score was compared between the low and high Drp1 subsets. Low expression:Remmele score ≤ 6, high expression:Remmele score > 6. Statistical significance was analyzed with chi-square test. ***P < 0.001. M Patients with different expression patterns of ROCK2 and Drp1 had different prognoses. The statistical significance was analyzed with log-rank test. The accuracy of the prognostic value of different expression patterns was evaluated with HR. N–P The correlation between the Drp1 expression and overall survival in patients with pCCA (n = 105, n (low expression) = 67, n (high expression)=38), dCCA (n = 90, n (low expression = 43), n (high expression) = 47), and iCCA(n = 38, n (low expression=26),n(high expression)=12)). Low expression:Remmele score ≤ 6, high expression:Remmele score > 6. The statistical significance was analyzed with log-rank test. The accuracy of the prognostic value of different expression patterns was evaluated with HR.