Figure 6: Chemokine gene expression with varying stiffness.

(a) Quantitative PCR (qPCR) of chemokine genes in the whole skin of P9 mice. Data represent mean±s.d. of six biological replicates assayed in triplicate. **P<0.01 and ***P<0.001 as compared with WT. ^##P<0.01 as compared with Snail Tg. (b) Cultured dermal fibroblasts were treated with conditioned media from Snail Tg keratinocytes (Snail CM) or WT keratinocytes (WT CM) for 18 h. Chemokine gene expression of treated fibroblasts was determined by qPCR. ***P<0.001 as compared with WT CM. (c) Dermal fibroblasts were plated on collagen-coated polyacrylamide hydrogels of varying stiffness (soft (2 kPa) and hard (5 kPa) gel). Cells were incubated with WT CM or Snail CM for 18 h with or without the presence of TGF-β receptor inhibitor SB-431542 at the concentration of 10 μM. Chemokine gene expression in treated fibroblasts was determined by qPCR. (d) Dermal fibroblasts were treated as in c with or without the presence of FAK inhibitor, PF 573228, at the concentration of 20 nM (FAK-I). Chemokine gene expression was determined by qPCR. (e) Dermal fibroblasts were transfected with a mock or CD2-FAK vector were treated as in d. Chemokine gene expression was determined by qPCR. Unless otherwise noted, data represent mean±s.d. of six samples, where *P<0.05, **P<0.01 and ***P<0.001. (d) ^##P<0.01, and ^###P<0.001 as compared with Soft gel+Snail CM (vehicle). ^†P<0.001 as compared with Hard gel+Snail CM. (e) ^#P<0.05, and ^##P<0.01 as compared with Soft gel+Snail CM (mock). (a–e) Statistical analyses were performed with Student's t-test.