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. 2023 Sep 6;12(18):2220. doi: 10.3390/cells12182220

Figure 3.

Figure 3

NOX4 overexpression partially restored CB-ECFC angiogenic function in hypoxia via activation of Nrf2. CB-ECFCs were electroporated with either an EV or NOX4 OE plasmid for 48 h and (A) protein was collected at 72 h for confirmation of NOX4 OE or incubated under hypoxia or normoxia for a further 48 h prior to quantification of (B) tube area using Matrigel tubulogenesis assay or (C) Nrf2 protein expression using Western blot with normalisation to β-actin as reference control. CB-ECFCs were treated with sulforaphane (SFN; 4 µmol/L for 4 h) prior to (D) visualisation of Nrf2 activation by immunocytochemistry, and (E) quantification of tube area using Matrigel tubulogenesis assay after further incubation under hypoxia for 48 h. Representative blots are shown from a single clone for each group. 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 the relevant EV/DMSO control, unpaired Student’s t-test or one-way ANOVA with Bonferroni post-hoc testing.