Figure 4.

Effect of osmotic pressure on matrix protease production of CAV1 CRISPR‐Cas9 knockout (KO) U251 cells. Cells were subjected to normo‐ (285 mOsmol/kg) or hyper (360 mOsmol/kg)‐osmolar medium. (A) Conditioned media of control and CAV1 KO cells exposed to osmotic stress were analysed by casein‐plasminogen zymography. Densitometric quantitation of the 47 and 51 kD bands corresponding to uPA was carried out. (B) Gelatin zymography and densitometric quantitation of MMP‐2 and MMP‐9 produced in the conditioned medium of control and CAV1 KO cells after 48 h of osmotic stress. (C) Effect of osmotic stress on uPA, MMP‐2 and MMP‐9 mRNA expression. The results are the average of n = 4 KO cell pools. Results are expressed as mean ± SEM of n = 3 independent experiments, *P < .05, **P < .01, ***P < .001, ****P < .0001, two‐way ANOVA analysis with Tukey's multiple comparisons test