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. 2021 Dec 6;24:87–100. doi: 10.1016/j.omto.2021.12.002

Figure 5.

Figure 5

SNX5 inhibits ccRCC cell stemness and CD44 endosomal trafficking

(A) The expression of CD44, Oct4, and SNX5 was detected by qRT-PCR in SNX5-overexpressing 786-O and 769-P cells. (B) The expression of CD44, Oct4, and SNX5 was detected by qRT-PCR in SNX5-knockdown 769-P and Caki-1 cells. Western blot analysis of CD44, Oct4, and SNX5 in SNX5-overexpressing cells (C) and SNX5 knockdown cells (D). (E) Western blot analysis of CD44 and SNX5 in SNX5-overexpressing tumor tissue and SNX5 knockdown tumor tissue. (F) CD44 on the cell surface in SNX5-overexpressing and SNX5 knockdown 769-P cells was assessed by flow cytometry. (G) SNX5-overexpressing 786-O and 769-P cells were applied to immunofluorescence staining. (H) IHC shows the expression of CD44 in SNX5-overexpressing tumor tissue and knockdown SNX5 tumor tissue. (I) Cells were surface-labeled with cleavable biotin at 4°C, left alone to allow internalization or recycling, and examined by western blotting. (J) Quantification of internalized CD44 in SNX5 overexpressing cells from (I). (K) Quantification of recycled CD44 in SNX5 overexpressing cells from (I). ∗p < 0.05; ∗∗p < 0.01.