Figure 6.

Inhibition of PLAC8 expression enhanced CB-ECFC NOX4-mediated angiogenic signalling in hypoxia. CB-ECFCs were transfected with either PLAC8-targeting siRNA or non-targeting SCR control for 24 h prior to culture in hypoxia or normoxia for 48 h. (A) Confirmation of PLAC8 KD by Western blot with normalisation to β-actin as reference control. (B) Tube area confirmed by Matrigel tubulogenesis assay. (C–E) Protein expression of NOX4, Nrf2 and eNOS by Western blot with normalisation to β-actin as reference control. Production of (F) superoxide confirmed using DHE staining and (G) hydrogen peroxide using a commercial assay with Image-J quantification. For scatter plots, data were mean ± SEM; n = 9 combined from three CB-ECFC clones; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus relevant SCR control, unpaired Student’s t-test or one-way ANOVA with Bonferroni post-hoc testing.