FIGURE 2.

Fibronectin matrix turnover and degradation are impaired in preeclampsia. (A) Western blot for FN in the matrix-soluble and -insoluble protein fractions from PTC (n = 3) and E-PE (n = 3) placental tissue (left panel) or TC and PE pMSCs (right panel) following sodium deoxycholate extraction. Representative of two separate pMSC isolations. (B) FN matrix turnover in TC and PE pMSCs, normalized to Actin (ACTB) over a time course of 24 h upon exposure to cycloheximide (CHX). n = 3 separate pMSC isolations. (C) Western blots (left panel) and corresponding densitometry (right panel) for FN in the lysosomal fraction isolated from PTC and PE placentae. The lysosomal marker, LAMP1, served as loading control. n = 7 separate placentae per group. (D) (left panel) IF for FN (green) and LAMP1 (red) in TC pMSCs exposed to NH₄Cl, compared to PE pMSCs. Nuclei were counter-stained with DAPI. Scale bars at 63 × magnification represent 7 μM. Representative of three separate pMSC isolations. (D) (right panel) Western blot for FN in TC and PE pMSCs exposed to 10 μM NH₄Cl for 24 h; − = H₂O Vehicle, + = 10 μM NH₄Cl. n = 3 separate pMSC isolations. (E) Western blot for FN in TC pMSCs exposed to 100 nM Bafilomycin for 24 h and in PE pMSCs; V = DMSO Vehicle, B = 100 nM Bafilomycin. (F) Western blot and corresponding densitometry for MAP1S in PTC and PE placental tissue. n = 6 separate placentae per group. Data are presented as mean ± SEM. *P ≤ 0.05 (unpaired Student’s t-test).