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. 2023 Apr 3;55(4):779–793. doi: 10.1038/s12276-023-00961-x

Fig. 8. SAMHD1 activates endosomal FAK signaling.

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.