Figure 3

Figure 3

TMEM184A siRNA KD decreases post-translational levels of VE-cad in the membrane and in cytoplasmic vesicles. (A) Comparisons of VE-cad (green) and TMEM CTD staining (red) in control siRNA (Con siRNA) and siTMEM BAOECs. Reduced levels of VE-cad and TMEM are highlighted in merged, white box, zoom, comparisons. Scale 20 μm. (B) Violin plots of corrected total cell fluorescence (CTCF) values of VE-cad and TMEM184A across Con siRNA (dark green and dark red violins) and siTMEM cells (light green and light red violins) in sub-confluent and confluent BAOECs across three independent experiments, n = 179 in sub-confluent condition, n = 193 in confluent condition, *** p < 0.0001. (C) WB of Con siRNA and siTMEM subcellular fractionation lysates comparing Cyt (supernatant) and Mem (pellet) fractions. Cyt fractions show full length VE-cad (130 kDa) and Actin loading control (42 kDa) with increased degradation in the siTMEM lane. In Mem fractions, full length VE-cad and Actin loading control bands are compared in one independent experiment. (D) Quantification of WB of subcellular fractionation densities of VE-cad normalized to Actin for full length VE-cad, VE-cad degradation products, and membrane VE-cad bands from one representative blot.