FIGURE 3.
The oxygen sensor, JMJD6, is required for fibronectin matrix formation. (A) Western blot and corresponding densitometry for FN in the first trimester (6–8 weeks, 9–12 weeks, and TC pMSCs). n = 3 separate pMSC isolations. (B) (left panel) Western blot and densitometry for JMJD6 and FN in TC pMSCs upon JMJD6 siRNA knockdown (siJMJD6). n = 3 separate pMSC isolations. (B) (right panel) IF for FN (green) and JMJD6 (red) following siJMJD6 in TC pMSCs. Nuclei were counter-stained with DAPI. Scale bars at 63 × magnification represent 7 μM. Representative of three separate pMSC isolations. (C) Western blot and corresponding densitometry for FN and JMJD6 upon JMJD6 siRNA knockdown (siJMJD6) in concert with NH4Cl exposure of TC pMSCs. n = 3 separate pMSC isolations. (D) Western blots and associated densitometry for JMJD6 and FN in TC pMSCs overexpressing (OE) WT JMJD6 (left panel) or mutated JMJD6 constructs (Mut JmjC; right panel). EV, vehicle control. n = 3 separate pMSC isolations, Actin (ACTB) served as loading control. Data are presented as mean ± SEM. *P ≤ 0.05 **P ≤ 0.01 (unpaired Student’s t-test; one-way ANOVA and Bonferroni post-test).