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. 2018 Jul 17;19(9):e45253. doi: 10.15252/embr.201745253

Figure EV3. PAR‐3 regulates EC Golgi reorientation toward flow in vitro .

Figure EV3

  • A
    Representative images of flow chamber‐cultured ECs transfected with control (Scrambled) or PARD3‐specific siRNA (siPAR‐3#1) and exposed to the indicated value of shear stress for 60 min. The cells were stained for nuclei (DAPI, blue) and Golgi apparatus (GOLPH4, red). Black arrows indicate corresponding axial polarity vectors. Flow direction is indicated on the right. Scale bar, 35 μm.
  • B, C
    Axial polarity of ECs treated with control (Scrambled) or PARD3‐specific siRNAs (siPAR‐3#1, B; or siPAR‐3#2, C) in response to 12, 18, and 30 dyn/cm2 laminar flow for the indicated time.
  • C
    Western blotting of EC lysates treated with 18 dyn/cm2 laminar flow for indicated times, with 1 μM of 6BIO (+) or control (−) containing growth medium. Upper panels show the blot of acetylated α‐tubulin (Ac α‐tubulin), and lower panels show total tubulin. Quantitation of relative intensity of Ac α‐tubulin is shown.
Data information: In (B and C), data are means ± SEM (n = 3 independent experiments and n = 100 cells for each experiment). In (D), data are means ± SEM (n = 3 experiments). Statistical significance (*P < 0.05; **P < 0.01) was evaluated with two‐way ANOVA and Bonferroni multiple comparisons post hoc analysis.