RT-qPCR analysis. HeLa cells were infected with JUP-specific shRNA lentiviral particles, total RNA was extracted from wild-type and knockdown cells, RT-qPCR was performed using gene-specific primers. ∆Ct (CtKD-CtWT) was used to calculate mRNA reduction (%) between wild-type and knockdown cells using the following formula: (1-1/2∆Ct) x 100%.

Western blotting analysis using anti-alpha smooth muscle Actin antibody (Cat#67172). alpha smooth muscle Actin antibody expression in wild type (WT) and alpha smooth muscle Actin antibody shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with anti-alpha smooth muscle Actin antibody (Cat#67172, 1:10,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).