Journal: bioRxiv

Article Title: Pancreatic cancer-associated organ dysfunction promotes muscle autophagy and contributes to peripheral tissue wasting

doi: 10.64898/2026.02.27.708635

Figure Lengend Snippet: a. Schematic showing experimental approach to disrupt autophagy in the muscle of mice with genetically engineered KP −/− F PDAC tumors. b. Western blot assessing Atg7 protein in muscle tissue from 6-week-old control ( Pdx-1-P2A-FlpO; Trp53 Frt/Frt ; Ckm-Cre ) or Atg7 muscle knock-out mice ( Ckm-Cre ; Atg7 fl/fl : referred to as Atg7 mKO), without or with ( Pdx-1-P2A-FlpO; Kras FSF-G12D/+ ; Trp53 Frt/Frt ) KP −/− F PDAC as indicated ( n = 3). Also shown is western blot for Atg7 in pancreas tissue from control and Atg7 mKO mice with KP −/− F PDAC ( n = 5). c. Western blot for Atg7, Atg5, p62, Lc3B-I, and Lc3B-II in gastrocnemius muscle from 6-week-old control and Atg7 mKO mice with KP −/− F PDAC ( n = 2). Vinculin is blotted as a loading control. d. Gastrocnemius, quadriceps, and soleus skeletal muscle weights normalized to whole-body weights from 6-week-old control or Atg7 mKO mice, without or with KP −/− F PDAC as indicated ( n = 14 control, n = 12 Atg7 mKO, n = 16 KP −/− F PDAC, and n = 11 Atg7 mKO KP −/− F PDAC). e. H&E staining of gastrocnemius muscle from 6-week-old control or Atg7 mKO mice, without or with KP −/− F PDAC as indicated. f. Myofiber area quantification from histology shown in e. ( n = 230 control, n = 166 Atg7 mKO, n = 477 KP −/− F PDAC, and n = 186 Atg7 mKO KP −/− F PDAC from n = 4 mice for each group). g-l. Whole-body weight ( n = 14 control, n = 14 Atg7 mKO, n = 16 KP −/− F PDAC, and n = 14 Atg7 mKO KP −/− F PDAC, 6-week-old) (g), food intake ( n = 5) (h), blood glucose ( n = 9) (i), plasma insulin ( n = 7 control, n = 6 Atg7 mKO, n = 5 PDAC, and n = 5 Atg7 mKO KP −/− F PDAC) (j), liver weight ( n = 11 control, n = 12 Atg7 mKO, n = 15 KP −/− F PDAC, and n = 11 Atg7 mKO KP −/− F PDAC) (k), and perigonadal and subcutaneous ATs weights ( n = 14 control, n = 12 Atg7 mKO, n = 16 KP −/− F PDAC, and n = 11 Atg7 mKO KP −/− F PDAC) (l) normalized to whole-body weights of 6-week-old control or Atg7 mKO mice, without or with KP −/− F PDAC as indicated. m. H&E staining of perigonadal AT from control and Atg7 mKO mice with KP −/− F PDAC. n. Pancreas tissue weights normalized to whole-body weights from 6-week-old control or Atg7 mKO mice, without or with KP −/− F PDAC as indicated ( n = 11 control, n = 12 Atg7 mKO, n = 11 KP −/− F PDAC, and n = 12 Atg7 mKO KP −/− F PDAC). o. Gross images of pancreas from 6-week-old control and Atg7 mKO mice with KP −/− F PDAC as indicated ( n = 2). p. H&E staining of pancreas tissue from 6-week-old control or Atg7 mKO mice, without or with KP −/− F PDAC as indicated. q. Survival of control mice with KP −/− F PDAC ( Pdx-1-P2A-FlpO; Kras FSF-G12D/+ ; Trp53 Frt/Frt , referred to here as KP −/− F PDAC), or the same mice with a Ckm-Cre and one ( Atg7 fl/+ ) or two ( Atg7 fl/fl ) Atg7 fl alleles as indicated ( n = 43 PDAC, n = 50 Atg7 fl/+ , and n = 82 Atg7 fl/fl ). r. Survival of control or Atg7 mKO mice following orthotopic implantation of murine KPC PDAC cells into pancreas ( n = 6). s. Survival analysis of control mice and mice with muscle-specific loss of function of Atrogin-1 ( Atrogin-1 fl/fl ; Ckm-Cre: Atrogin-1 mLOF) following orthotopic pancreatic implantation of KPC PDAC cells ( n = 6). t. Survival analysis of control mice and mice with combined muscle-specific Atrogin-1 loss of function and Atg7 deletion ( Atrogin-1 mLOF; Atg7 mKO) following orthotopic KPC PDAC implantation ( n = 6). Both male and female mice were used in experiments. Statistical analyses were performed using one-way ANOVA with Tukey’s post hoc test for data in panels c-j and m-n. P-values on survival experiments were calculated with Gehan-Breslow-Wilcoxon test. Data are presented as mean ± S.D., and n denotes the number of mice analyzed. Scale bars: 100 μm.

Article Snippet: The following mouse strains were studied in here: C57BL/6J mouse (The Jackson Laboratory, 000664), Nu/J mouse (The Jackson Laboratory, 002019), Ckm-Cre (The Jackson Laboratory, 006475), the conditional LysoTag mouse commonly known as Conditional LysoTag (The Jackson Laboratory, 035401), the conditional Atrogin-1 fl/fl (Taconic, 9569), conditional Atg7 fl/fl mouse (provided by Dr. Eileen White from Rutgers University, New Brunswick, NJ-USA).

Techniques: Western Blot, Control, Knock-Out, Staining, Clinical Proteomics