Research Paper Volume 15, Issue 6 pp 1808—1832
Cellular senescence and disrupted proteostasis induced by myotube atrophy are prevented with low-dose metformin and leucine cocktail
Figure 4. MET+LEU maintained proteostasis during SD. (A) Volcano plot of changed transcripts with MET+LEU vs. SD with a significance cut of -log10Adjusted p-value of 2, highlighting ribosomal subunit genes. (B) Myosin heavy chain genes from RNA-sequencing in 2% HS, SD, and MET+LEU. (C) Pathways altered with MET+LEU vs. SD related to proteostasis, numbers are representative of normalized enrichment scores (NES). (D) Puromycin incorporation after 2-days (D2), 3-days (D3), and 4-days (D4) of SD and representative western blot. (E) Proteasome activity assay results represented as relative fluorescent units, (E inset) calculated proteasomal activity rate. (F) Experimental schematic of 16.8 μM MG-132, or 0.1 + 0.5 μM MET+LEU treatment with SD. (G) Myotube area during 4 days SD, MET+LEU, or MG-132. ****p < 0.0001, **p < 0.01, *p < 0.05. N = 6/group for (A–D), N = 4/group for (E and F).