Fig. 8: SAMHD1 activates endosomal FAK signaling. | Experimental & Molecular Medicine

Fig. 8: SAMHD1 activates endosomal FAK signaling.

From: SAMHD1-induced endosomal FAK signaling promotes human renal clear cell carcinoma metastasis by activating Rac1-mediated lamellipodia protrusion

Fig. 8

a, b Western blot analysis of phospho-FAK (Y397) and total FAK levels in SAMHD1-knockdown ACHN and SN12C cells or SAMHD1-overexpressed Caki-1 and Caki-2 cells. c Western blot analysis of phosphorylated FAK (Y397) and total FAK expression in SAMHD1-overexpressed Caki-1 cells treated with 80 μM Dynasore over 24 h. d SAMHD1-overexpressed Caki-1 cells were treated with 10 μM PF573228 for 24 h and immunolabeled for EEA1. The nuclei were stained with DAPI. Scale bar, 20 μm. e Graphs show the number of EEA1 vesicles per single cell (n = 55 cells). ***p < 0.001. f IP assay was performed using Flag-tagged beads in Flag-SAMHD1-overexpressing Caki-1 cell lysates treated with 10 μM PF573228 over 24 h. g Western blot analysis of phosphorylated FAK (Y397) and phosphorylated cortactin (Y421) expression in SAMHD1-overexpressed Caki-1 cells treated with 10 μM PF573228 over 24 h. h, i Wound-healing assay of cell migration ability in SAMHD1-knockdown SN12C cells or SAMHD1-overexpressed Caki-1 cells treated with 10 μM PF573228. The relative wound-healing area was normalized to that of the controls. Data are presented as the mean ± SEM. The experiments were performed at least three times. *p < 0.05, ***p < 0.001. j IHC of SAMHD1 and p-FAK (Y397) expression in tissues of patients with ccRCC. Scale bar, 50 μm. k Areas with positive IHC staining were measured and are presented in the graph. Data are presented as the mean ± SD. *p < 0.05. l SAMHD1 promotes ccRCC metastasis through endosomal FAK signaling activation.

Back to article page