Figure 7.
Talin-1 in sEVs does not regulate the binding affinity of integrins for laminin. (A) Western blot analysis of PC3 and BxPC3 cells after talin-1 KD by siRNA. (B and C) Cell spreading assay of WT and talin-1 (Tln1) KD PC3 cells (B) and BxPC3 cells (C) on glass coated with ECM components: fibronectin (FN), laminin (LN), or collagen typeⅠ (COL1). Cells were observed after 2 h of incubation, and cell areas were quantified (n = 35 cells). (D) Western blot analysis of PC3 cell–derived sEVs after talin-1 KD by siRNA or overexpression of talin-1. (E–G) The fluorescence images (E) and the numbers of PC3-sEVs attached to glass coated with laminin before and after (F) talin-1 KD or (G) overexpression of talin-1 (n = 14 images). Halo7-integrin β1 in sEVs was labeled with SF650T. (H–J) The fluorescence images (H) and the numbers of CD63-labeled sEVs attached to glass coated with laminin before and after (I) talin-1 KD and (J) overexpression of talin-1 (n = 16 images). (K and L) The numbers of EV–CD63Halo7-SF650T particles attached to glass coated with collagen type I before and after (K) talin-1 KD or (L) overexpression of talin-1 (n = 21 images). (M) Western blot analysis of BxPC3 cell–derived sEVs after talin-1 KD by siRNA. (N–P) Fluorescence images (N) and the numbers of sEVs-BxPC3 bound to laminin (O) or collagen type I (P) on glass before and after talin-1 KD (n = 7 images). The membranes of sEVs were stained with Exosparkler DeepRed. (Q) Western blot analysis of the phosphorylation of Ser425 on talin-1 in PC3 cells and sEVs. Roscovitine: an inhibitor of CDK5 that phosphorylates Ser425 of talin-1. (R) Western blot analysis of kindlin-2 in PC3 cells and PC3-derived sEVs before and after kindlin-2 KD and talin-1 KD. Data are presented as the mean ± SE. n.s., nonsignificant difference; *P < 0.05; **P < 0.01; ***P < 0.001 according to Welch’s t test (two-sided). Source data are available for this figure: SourceData F7.

Talin-1 in sEVs does not regulate the binding affinity of integrins for laminin. (A) Western blot analysis of PC3 and BxPC3 cells after talin-1 KD by siRNA. (B and C) Cell spreading assay of WT and talin-1 (Tln1) KD PC3 cells (B) and BxPC3 cells (C) on glass coated with ECM components: fibronectin (FN), laminin (LN), or collagen typeⅠ (COL1). Cells were observed after 2 h of incubation, and cell areas were quantified (n = 35 cells). (D) Western blot analysis of PC3 cell–derived sEVs after talin-1 KD by siRNA or overexpression of talin-1. (E–G) The fluorescence images (E) and the numbers of PC3-sEVs attached to glass coated with laminin before and after (F) talin-1 KD or (G) overexpression of talin-1 (n = 14 images). Halo7-integrin β1 in sEVs was labeled with SF650T. (H–J) The fluorescence images (H) and the numbers of CD63-labeled sEVs attached to glass coated with laminin before and after (I) talin-1 KD and (J) overexpression of talin-1 (n = 16 images). (K and L) The numbers of EV–CD63Halo7-SF650T particles attached to glass coated with collagen type I before and after (K) talin-1 KD or (L) overexpression of talin-1 (n = 21 images). (M) Western blot analysis of BxPC3 cell–derived sEVs after talin-1 KD by siRNA. (N–P) Fluorescence images (N) and the numbers of sEVs-BxPC3 bound to laminin (O) or collagen type I (P) on glass before and after talin-1 KD (n = 7 images). The membranes of sEVs were stained with Exosparkler DeepRed. (Q) Western blot analysis of the phosphorylation of Ser425 on talin-1 in PC3 cells and sEVs. Roscovitine: an inhibitor of CDK5 that phosphorylates Ser425 of talin-1. (R) Western blot analysis of kindlin-2 in PC3 cells and PC3-derived sEVs before and after kindlin-2 KD and talin-1 KD. Data are presented as the mean ± SE. n.s., nonsignificant difference; *P < 0.05; **P < 0.01; ***P < 0.001 according to Welch’s t test (two-sided). Source data are available for this figure: SourceData F7.

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