Figure 1.

LPA induces mTORC2 signaling-associated mSin1 in mesenchymal cells. A, nonfibrotic MCs were pretreated with the mTORC inhibitor AZD8055 (250 nm for 30 min) and then exposed to LPA (10 μm for 6 h). Protein expression of the total and the phosphorylated forms of AKT at Thr-308 and Ser-473 were measured by immunoblotting. B, cell lysates isolated from normal nonfibrotic MCs were immunoblotted for antibodies against mTORC2-associated mSin1 and rictor, mTORC1-associated raptor, and GAPDH (loading control (C)). C, quantitative densitometry of the mTORC2 binding partner mSin1 was analyzed at 6 and 16 h in response to LPA. Values are means ± S.D., n = 4–6. *, p < 0.05; **, p < 0.01. D, nonfibrotic MCs were exposed to TGF-β (2 ng/ml) or IL-13 (10 ng/ml) for 16 h. Lysates were immunoblotted against mSin1. Quantitative densitometry was normalized using GAPDH (loading control). Values are means ± S.D., n = 9. *, p < 0.05; **, p < 0.01. Statistics: one-way ANOVA; post hoc: Sidak (C and D).