1. KLF4 regulates the EndMT switch in CCM1 KO ECs
KLF4 regulates the EndMT switch in CCM1 KO ECs A–DCultured lung‐derived WT and CCM1 KO ECs were lentivirally transduced with shRNA directed to either Klf4 (shKLF4) or control sequence (shCTRL). (A) qRT–PCR of mesenchymal (Fsp1, Id1), stem cell‐like (Sca1), and endothelial markers (VE‐cadherin and Claudin5) in WT shCTRL, WT shKLF4, CCM1 KO shCTRL, and CCM1 KO shKLF4 ECs. Data are mean ± SD (n = 3). Fold difference in gene expression is relative to WT shCTRL ECs. A two‐tailed unpaired t‐test was performed. Klf4: ****P <  , ***P = 0.0008, **P = 0.007; Fsp1: ***P = 0.0001, ****P = 3.9E‐05, ###P = 0.0002; Sca1: ***P = 0.0001, ****P <  ; Id1: ***P = 0.0001; Ve‐cadherin: ***P = 0.0001; Claudin5: ****P = 1.37E‐05. (B) WB of EndMT markers in WT shCTRL, WT shKLF4, CCM1 KO shCTRL, and CCM1 KO shKLF4 ECs. Vinculin is the loading control. These data are representative of three independent observations. (C) Proliferation rate of WT shCTRL, WTshKLF4, CCM1 KO shCTRL, and CCM1 KO shKLF4 ECs cultured for 5 days. Columns represent mean ± SD (n = 8). A two‐tailed unpaired t‐test was performed. 3 days of culture: ***P = 0.0001, **P = 0.0045, ##P = 0.0017; 5 days of culture: ***P = 0.0001, **P = 0.0029, ##P =  (D) Migration rate measured in a wound assay of WT shCTRL, CCM1 KO shCTRL, and CCM1 KO shKLF4 ECs. Mean ± SD is shown (n = 6). A two‐tailed unpaired t‐test was performed. ***P = 0.0004, ###P =  ECultured lung‐derived WT ECs were lentivirally transduced with a full‐length murine Klf4 (LentiKLF4) or empty vector (Mock). qRT–PCR (left panel) and WB (right panel) of EndMT markers in Mock and LentiKLF4 ECs. qRT–PCR data are mean ± SD (n = 3) and the fold changes are relative to Mock ECs. A two‐tailed unpaired t‐test was performed. **P = 0.002, ***P = 0.0007, ###P =  WB results are representative of three independent observations. Tubulin is the loading control.Source data are available online for this figure.
Roberto Cuttano et al. EMBO Mol Med. 2015;emmm
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