Figure 3
From: Matrix metalloproteinase-12 is an essential mediator of acute and chronic arterial stiffening
Similar effects of ECM stiffness on wild-type and MMP12-null SMCs in culture.
(A) Mouse (n = 4) and human (n = 5) vascular SMCs were serum-starved for 48 h and then seeded on low and high stiffness fibronectin-coated acrylamide hydrogels with 10% FBS for 24 h. The bar graph shows mean + SD with the level of MMP12 mRNA in the low stiffness hydrogels set to 1.0. (B–D) Wild-type and MMP12-null SMCs were serum-starved and incubated on low (L)-, medium (M), or high- (H)-stiffness hydrogels (2–4, 10–12, and 20–25 kPa, respectively) with 10% FBS for 24 hr. (B) Intracellular stiffness was determined by AFM. The bar graph shows mean + SE of 4 independent experiments with 10 cells analyzed per experiment. (C) Cells were co-stained with phalloidin (red) and anti-paxillin (green). Replicate coverslips were stained with anti-FAKPY397 (red); n = 4 independent experiments with 5 cells analyzed per experiment. (D) S-phase entry was determined by EdU incorporation after a 72-hr incubation in 10% FBS. The bar graph shows mean + SD; n = 4. p values are from two-tailed Mann-Whitney tests.
