Maintaining resting cardiac fibroblasts in vitro by disrupting mechanotransduction
Fig 6
Reversed cardiac fibroblasts re-differentiate on plastic.
A) Schematic illustration of re-differentiation protocol. Cardiac fibroblasts were cultured on plastic for 6 days (d) and then treated with blockers of ROCK (Y27) and TGFβRI (SB) for 3d, where after blockers were removed and cells transferred to soft gels for 4d. B) Immunostaining for smooth muscle α-actin (SMA; green). Nuclei are stained with DAPI (blue). Scale bar 100μm. C) mRNA expression of actin alpha 2 smooth muscle (Acta2), connective tissue growth factor (Ctgf), collagen (Col) 1a1, Col1a2, Col3a1, lysyl oxidase (Lox) and periostin (Postn) relative to reversed cells (grey bars), following re-differentiation on plastic (black bars) or soft gels (white bars). Significant differences were determined by Student’s t-test with Holm-Sidak correction for multiple comparisons. N = 3.