Fig. 6.

Fzd5 knockdown led to increased expression of vascular regression-associated genes Flt1 and Angpt2, independent of the non-canonical Wnt/Ca²⁺ and PCP pathways. a QPCR results of expression levels of NFAT target gene Dscr1 in the different conditions 72 h post-transfection and in response to ionophore A23187 (10 µM)-induced Ca²⁺ flux with and without NFAT inhibitor Cyclosporin A (CsA) (1 µM). N = 5, *P < 0.05 compared to control and siSHAM condition, and DMSO-treated and CsA + A23187-treated ECs, respectively. b Angpt2 and Flt1 mRNA expression levels in HUVECs in response to CsA, supplemented 48 h post-transfection. N = 4, *P < 0.05 compared to control and siSHAM condition (two-way ANOVA followed by Bonferroni post hoc test). c Representative Western blot of total JNK, phospho-JNK, and β-actin levels at different time points post-transfection. d Quantified results of JNK and phospho-JNK Western blot. Shown are individual (phospho) JNK isoform (p46 and p54) levels relative to β-actin loading control. N = 6, *P < 0.05 compared to control and siSHAM condition within one time comparison (24, 48 or 72 h). e Western blot of total JNK, phospho-JNK, and β-actin levels in response to different concentrations of JNK inhibitor SP600125 after 1 h. f QPCR analysis showing the effect of SP600125, supplemented 48 h post-transfection, on Flt1 and Angpt2 mRNA levels in the different conditions. N = 4, *P < 0.05 compared to control and siSHAM condition, ^#P < 0.05 as indicated in graph (two-way ANOVA followed by Bonferroni post hoc test)