Fig. 5: Silencing of PIM3 dismantles cadherin-5 and catenins from endothelial cell-cell junctions.

a shPIM3 or control (shScr) silenced human umbilical vein endothelial cells (HUVECs) and dermal microvascular endothelial cells (BECs) were stained for vascular endothelial cadherin (CDH5) and F-actin. Nuclei were stained using DAPI. Relative CDH5 signal intensity (b) and area (c) (normalized to number of nuclei) in HUVEC (n = 3 independent experiments for shScr, n = 6 independent experiments for shPIM3) and in BEC (n = 3 for shScr, n = 5 for shPIM3). Western blot (d) and quantification (e) of CDH5 in HUVECs treated as in (a). n = 3 independent experiments for shScr, n = 6 for shPIM3. f shPIM3 or shScr silenced BECs were stained for α- and β-catenin (CTNNA1 and CTNNB1) and HUVECs for δ-catenin (CTNND1) and F-actin. Nuclei were stained using DAPI. Relative α-catenin (g, h), β-catenin (i, j) and δ-catenin (k, l) signal intensities (per field) and area (normalized to number of nuclei) relative to control (shScr). n = 3 for shScr, n = 6 for shPIM3 (g–j). n = 3 for shScr, n = 5 for shPIM3 (k, l). m Schematic representation of CDH5 and α-, β- and δ-catenin in adherens junctions created in Biorender.com. Two-tailed unpaired t-test (b, c, e, g–l). Data are presented as mean values ⁺/- SD. Data is pooled from independent experiments using two shPIM3 clones in b, c, e, g–l. Scale bars 50 μm (a, f); 25 μm in close-up images (a, f). Source data are provided as a Source Data file.