Figure 4. Klf4 overexpression prevents shear stress induced EndoMT.

(A,B) QPCR and Western Blot analysis, respectively, showing Klf4 overexpression in Tg737^orpk/orpk EC which were transfected with either LacZ (Tg737^o/o-LacZ, sham) or Klf4 (Tg737^o/o-Klf4) expression constructs. Transfection with Klf4 results in a 10 fold overexpression on mRNA level, and a 2.2 fold overexpression on protein level. (C) Images of Tg737^orpk/orpk-LacZ and Tg737^orpk/orpk-Klf4 cells under static conditions and under 0.5 Pa shear stress. Arrows indicate the direction of flow. Scale bars: 25 μm. (D) Q-PCR showing relative mRNA expression of CD31, αSMA, Pai1, Snai1, and Ncad in Tg737^orpk/orpk-LacZ and Tg737^orpk/orpk-Klf4 cells under 0.5 Pa shear stress. Expression is normalized to GAPDH and relative to static shams, as represented by the dashed line. (E) Western Blot analysis and quantification of CD31 protein in Tg737^orpk/orpk-LacZ and Tg737^orpk/orpk-Klf4 cells under static conditions and 0.5 Pa shear stress.