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. 2017 Jan 23;7:41173. doi: 10.1038/srep41173

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Copyright © 2017, The Author(s)

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(a) LARP6 phosphorylation changes in activation of HSCs. Primary rat HSCs were transduced by adenovirus expressing wt LARP6 on day 2 or on day 4 and analyzed by 2DGE on day 3 or on day 5 (CON, panels 1 and 2). The same cells were also treated with rapamycin on day 2 and day 4 and analyzed on day 3 and day 5 (panels 3 and 4). Panels 5 and 6; cells transduced with S348A/S409A mutant. (b) Increased AKT and mTORC1 signaling during HSCs activation. HSCs at day 3 and day 5 in culture were analyzed for expression of phospho-AKT and phospho-S6K. p-AKT: phopho-AKT. AKT: total AKT. p-S6K: phospho-S6K. S6K: total S6K. ACT: β-actin, loading control. (c) Dominant negative effect of S348A/S409A in HSCs. HSCs were transduced on day 3 with wt LARP6 and S348A/S409A mutant and on day 5 cellular (CELL) and medium (MED) level of COL1A1 was analyzed by Western blot. Loading controls: β-actin (ACT) and fibronectin (FIB). HA-LARP6, expression of the transduced proteins. (d) Rapamycin reduces expression of collagen by HSCs. HSCs cultured for 5 days were treated with or without RAPA for 2 h and COL1A1 polypeptide was analyzed intracellularly (CELL) or in the medium (MED).