Skip to main content
. 2017 Jan 23;7:41173. doi: 10.1038/srep41173

Figure 2. Inhibition of mTORC1 reduces secretion of type I collagen.

Figure 2

(a) Intracellular collagen polypeptides in HLFs treated with RAPA. HLFs were treated with CON or RAPA for 24 h and intracellular (CELL) level of collagen α1(I) (COL1A1) and α2(I) (COL1A2) polypeptides was analyzed by Western blot. Loading control: β-actin (ACT). (b) Level of collagen polypeptides in the cellular medium. Collagen polypeptides in the medium (MED) of cells in (a) were analyzed by Western blot. Loading control: fibronectin (FIB). (c) Rapamycin does not change expression of collagen mRNAs. Total RNA from cells in (a) was analyzed for expression of COL1A1 and COL1A2 mRNA by real-time RT-PCR. Expression was normalized to the internal control β-actin mRNA and presented as mean ± SD (n = 3). (d) Change in pI of COL1A2 by rapamycin. COL1A2 from control HLFs or HLFs treated with RAPA for 24 h was analyzed by 2DGE. Arrow: isoelectric focusing of COL1A2 in control cells. (e) Time course of inhibition of type I collagen by rapamycin. HLFs were treated with RAPA for the indicated time periods and intracellular (CELL) level of collagen polypeptides was analyzed by Western blot. CON; untreated cells. (f) Time course of inhibition of collagen secretion. Collagen in the medium of cells in (e) was analyzed by Western blot.