Fig. 1
From: The TRPV4 channel links calcium influx to DDX3X activity and viral infectivity

TRPV4 interacts with DDX3X. a Co-immunoprecipitation (Co-IP) of TRPV4-V5(tag) and DDX3X-Myc(tag) in HEK293 cells. b Co-IP of endogenously expressed TRPV4 and DDX3X in Huh7 cells. Uncropped westerns are shown in Supplementary Fig. 3d–f. c Quantification of immunoprecipitated DDX3X normalized by DDX3X input and immunoprecipitated TRPV4. Mean ± SEM of three Co-IPs of heterologously expressed or endogenously expressed TRPV4 and DDX3X proteins. *P < 0.05 (P = 0.012) two-tailed Student’s t test for heterologously and P = 0.036 for endogenously expressed proteins when compared control with GSK conditions. d Confocal immunofluorescence images of TRPV4 (cyan) and DDX3X (magenta) in HeLa cells overexpressing TRPV4-V5 and DDX3X-Myc. Co-localized pixels are shown in white in the merge panels. Scale bar = 10 μm. e Plot profile analysis performed using ImageJ software on each image along the white line shown on the panels. f Pearson’s coefficient of the TRPV4 interaction with DDX3X under control conditions and upon channel activation with GSK1016790A. g FRET efficiency represented as the CFP increase during YFP photobleaching normalized to the initial CFP value measured in HeLa cells expressing TRPV4–CFP and DDX3X-YFP or free soluble YFP, and exposed to control condition or in the presence of 10 nM GSK1016790A. The number of cells analysed is indicated in each bar. Data are means ± SEM. ***P < 0.001 (P = 0.0008), two-tailed Student’s t test for f and **P < 0.01 one-way ANOVA followed by a Bonferroni post hoc test (P = 0.007) when comparing control TRPV4-CFP/free-YFP vs. any other condition (g). Bartlett’s test for equal variances reported no significant differences (P = 0.6)