Figure Legends
Figure 1 . Collagen deposition within skeletal muscle measured via Picrosirius Red staining during the progression of cancer cachexia in the A) plantaris (female) and B) tibialis anterior muscle (male).N of 4-5 per group. Different letters denote statistical significance at an alpha set a P ≤ 0.05. 1wk, 2wk, 3wk, 4wk, denotes 1, 2, 3, and 4 weeks of cancer progression, respectively; LT and HT denote low tumor and high tumor, respectively; PBS, phosphate-buffered saline. Different letters denote statistical significance at an alpha set a P ≤ 0.05.
Figure 2 . Collagen 1 and 3 mRNA abundance during the progression of cancer cachexia in females (left) and males (right). (A-B) Collagen 1 and 3 mRNA abundance in female tibialis anterior muscle. (C) Collagen 3:1 mRNA abundance female tibialis anterior muscle. (D-E) Collagen I and III mRNA abundance in male tibialis anterior muscle. (F) Collagen 3:1 mRNA abundance male tibialis anterior muscle. For females, N of 10-14 per group were used. For males, Nof 7-8 per group were used. 1wk, 2wk, 3wk, 4wk, denotes 1, 2, 3, and 4 weeks of cancer progression, respectively; LT and HT denote low tumor and high tumor, respectively; PBS, phosphate-buffered saline. Different letters denote statistical significance at an alpha set a P ≤ 0.05.
Figure 3. mRNA abundance of markers of extracellular matrix remodeling and turnover during the progression of cancer cachexia in females (left) and males (right). (A-B) MMP2 and MMP9 mRNA abundance in female tibialis anterior muscle. (C-D) MMP2 and MMP9 mRNA abundance in male tibialis anterior muscle. For females, N of 10-14 per group were used. For males, N of 7-8 per group were used. 1wk, 2wk, 3wk, 4wk, denotes 1, 2, 3, and 4 weeks of cancer progression, respectively; LT and HT denote low tumor and high tumor, respectively; PBS, phosphate-buffered saline. Different letters denote statistical significance at an alpha set a P ≤ 0.05.
Figure 4 . mRNA abundance of the TGF-β/SMAD signaling pathway components during the progression of cancer cachexia. (A-C) Relative mRNA abundance of TGF-β1, SMAD 2/3 in female tibialis anterior muscle. (D-F) Relative mRNA abundance of TGF-β1, SMAD 2/3 in male tibialis anterior muscle. For females, N of 10-14 per group were used. For males, N of 7-8 per group were used. 1wk, 2wk, 3wk, 4wk, denotes 1, 2, 3, and 4 weeks of cancer progression, respectively; LT and HT denote low tumor and high tumor, respectively; PBS, phosphate-buffered saline. Different letters denote statistical significance at an alpha set a P ≤ 0.05.
Figure 5. Effect of LCM treated media on C2C12 myotube size. (A) Representative images of C2C12 myotubes in media treated with vehicle or LCM. (B) Myotube diameter on C2C12 myotubes treated with either Vehicle (Veh) or LLC conditioned media (LCM). Different letters denote statistical significance at an alpha set a P ≤ 0.05.
Figure 6 . Effect of LCM treated media and co-culture on ECM regulation in C2C12 myotubes. (A-C) mRNA abundance of collagen1, 3, and the collagen 3 to 1 ratio. (D-F). mRNA abundance of ECM remodeling and turnover markers MMP2, 9, and TIMP-1. Different letters denote statistical significance at an alpha set a P ≤ 0.05.
Figure 7. Effect of LCM treated media and co-culture on SMAD mRNA abundance in C2C12 myotubes or 3T3 fibroblasts treated with vehicle (Veh) or LLC conditioned media (LCM) treated. Different letters denote statistical significance at an alpha set a P ≤ 0.05.