Fig. 5
From: The TRPV4 channel links calcium influx to DDX3X activity and viral infectivity

TRPV4 promotes nuclear export and translation of viral RNA. a Translation of viral RNA2 in yeast. Western blot analysis of protein 2A (top), protein PGK (middle) and qPCR quantification of viral RNA2 normalized to actin (lower) obtained in WT, ∆Yvc1 and ∆Yvc1 yeast transformed with hTRPV4. Differences in mean RNA2 values were not statistically significant (P > 0.05, n = 3) as determined by one-way ANOVA followed by a Bonferroni post hoc test when compared WT yeast with ∆Yvc1 or ∆Yvc1+hTRPV4 (P = 0.26 and P = 0.08, respectively). b Quantification of RNA2 translation, calculated as the ratio of corrected protein 2a/RNA2. c Representative images of HeLa cells transfected with Rev-mCherry, RRE-gRNA and MS2-YFP, and stained with DAPI. d Quantification of nuclear export of gRNA, calculated as the difference in the total/nuclear MS2-YFP ratio between HC067047 and control conditions in cells transfected with siControl, siTRPV4 and siDDX3X. Mean ( ± SEM) of 67 siControl cells, 46 siTRPV4 cells and 47 siDDX3X analyzed from two independent experiments. e Quantification of Gag-CFP signal expressed as a negative difference in the GAG-CFP intensity between HC067047 and control conditions in cells transfected with siControl, siTRPV4 and siDDX3X. Cell contour marked by a dotted line. Mean ± SEM of 67 siControl cells, 46 siTRPV4 cells and 47 siDDX3X analyzed from two independent experiments. The data are means ± SEM. *P < 0.05 and ***P < 0.001 when comparing WT with any other condition (b) or siControl versus any other condition (d, e) as determined by one-way ANOVA followed by a Bonferroni post hoc test. Bartlett’s test for equal variances reported significant differences in d and e (P = 0.01 and P = 0.0001, respectively). Scale bar = 10 μm