Extended Data Fig. 6: The role of TRPV4 in cell mechanotransduction in response to extracellular fluid viscosity.
From: Extracellular fluid viscosity enhances cell migration and cancer dissemination
a, (Left) Representative western blot of SC and shTRPV4 cells using two shRNA sequences. Image is representative of 2 independent biological replicas. (Right) Relative TRPV4 mRNA levels of SC and shTRPV4 cells. Data are normalized to the levels of SC cells, and represent the mean ± range from 2 experiments. b, Representative whole-cell TRPV4 current traces in SC (left) and shTRPV4 (right) cells exposed to 0.77 cP (pink) or 8 cP in the absence (blue) or presence (grey) of the TRPV4 inhibitor HC-067047 (HC). c, GCaMP6s activity in SC, shTRPV4 or shNHE1 cells at 0.77 or 8 cP. d,e, Confined migration speeds of SC and shTRPV4 (sequence 1) cells (d), or wild-type cells under GSK 2193874 (GSK2) or vehicle control treatment (e) at prescribed extracellular viscosities. Data are mean ± s.d. for n≥140 cells from 3 experiments. f, Effect of TRPV4 inhibition via GSK 2193874 (GSK2) in SUM159, HOS and U87 cells or TRPV4 knockdown (sequence 1) in brain metastatic MDA-MB-231 cells (BrM2) on confined migration speeds at the prescribed viscosities. Data are mean ± s.d. for n≥83 cells from ≥2 experiments. g, Representative whole-cell TRPV4 current traces in SC (left) and shNHE1 (right) cells exposed to 0.77 cP (pink) or 8 cP in the absence (blue) or presence (grey) of the TRPV4 inhibitor HC-067047 (HC). h, GCaMP6s activity in SC or shRNA β1-integrin (shITGB1) cells at the prescribed viscosities. Data are mean ± s.d. for n≥32 cells from 2 experiments. Tests performed: one-way ANOVA followed by Tukey’s multiple comparisons (f (only HOS)) and after log transformation of data (d,e), Kruskal-Wallis followed by Dunn’s multiple comparison (f (except HOS), h). For gel source data, see Supplementary Fig. 1. Cell model: MDA-MB-231 unless otherwise indicated.
