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. 2016 Oct 5;11(10):e0162636. doi: 10.1371/journal.pone.0162636

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© 2016 Biswas, Longmore

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 4 — SNAIL1-deleted cardiac fibroblasts have decreased collagen I mRNA expression and collagen matrix deposition. (A) Relative SNAIL1 mRNA level in isolated SNAIL1^f/f; ROSA-LSL-tdTomato cardiac fibroblasts treated with Adeno-LacZ (CTL) or Adeno-Cre (+ Cre), with and without TGFβ stimulation (2ng/ml for 2 hours), as determined by Q-PCR. SNAIL1 mRNA level in CTL unstimulated cells was arbitrarily set to 1. (B) Western blot analysis, with the indicated antibodies, of extracts from cardiac fibroblasts isolated from SNAIL1^f/f; ROSA-LSL-tdTomato mice and treated with Adeno-LacZ (CTL) or Adeno-Cre (+ Cre), with and without TGFβ stimulation (2 ng/ml for 2 hours). Col1α1 (C) and Col3α1 (D) mRNA levels at baseline and following TGFβ stimulation (2 ng/ml for 2 hours) in control (CTL) and SNAIL1^-/- (+ Cre) cardiac fibroblasts. Shown is a representative result of one of 4 independent experiments. (E) Collagen I immunofluorescence of cell free matrix deposited by control (CTL) and SNAIL1^-/- (+ Cre) cardiac fibroblasts. (n = 3). (F) Quantification of results in (E). (G) Detection of Collagen 1 by Sirius Red Staining of cell free matrix deposited by control (CTL) and SNAIL1-deleted (+ Cre) cardiac fibroblasts. (n = 4). (H) Birefringence imaging of Sirius red stained matrix produced by control (CTL) or SNAIL^-/- (+ Cre) cardiac fibroblasts. (I) Quantification of birefriengence imaging (n = 4, 20 images counted for each experiment). Scale bars: 50 microns.