Figure 3 from Tumor-Secreted ADAMTSL4 Activates Latent TGFβ1 to Drive Cancer Cachexia

Representative structural features are highlighted by arrows (white, PBS/control; red, cachectic condition; blue, ADAMTSL4 knockdown). <b>K,</b> Quantification of muscle fiber CSA (<i>n</i> = 3 per group, >500 fibers per mouse) and (<b>L</b>) fiber size distribution in TA. <b>M,</b> Selected GO terms from GSEA of GC muscle from C26-shCtrl vs. PBS-injected mice (see also Supplementary Data S3 and S4 for full GO term analysis results). <b>N,</b> Selected GO terms from GSEA of GC muscle from C26-shAdamtsl4 vs. C26-shCtrl tumor-bearing mice (see also Supplementary Data S5 and S6 for full GO term analysis results). <b>O,</b> Quantification of Western blot analysis of key proteins involved in proteasomal catabolism and autophagy, including p-FOXO3a at T32 and p-STAT3 at Y705. <b>P,</b> mRNA expression of proteasomal catabolic genes (<i>Fbxo32</i> and <i>Trim63</i>) and (<b>Q</b>) autophagy-related genes (<i>Becn1</i>, <i>Map1lc3</i>, and <i>Gabarapl1</i>) validated by qPCR. <b>R,</b> Quantification of Western blot analysis of autophagy marker LC3, showing LC3-I, LC3-II, and the LC3-II/LC3-I ratio.

Data are presented as the mean ± SEM. Each dot represents an individual biological replicate. Statistical analysis: two-tailed unpaired <i>t</i> test with Welch correction (<b>B</b> and <b>D</b>), one-way ANOVA with Tukey multiple-comparisons test (<b>C</b>, <b>F</b>, <b>G–I</b>, <b>K</b>, and <b>O–R</b>), two-way ANOVA with mixed-effects model (REML) followed by Tukey’s test (<b>E</b> and <b>G</b>), and GSEA enrichment scores calculated using weighted Kolmogorov–Smirnov statistics (<b>M</b> and <b>N</b>).

Significance is denoted as *, <i>P</i> < 0.05; **, <i>P</i> < 0.01; ***, <i>P</i> < 0.001; ****, <i>P</i> < 0.0001; ns, not significant. [<b>A,</b> Created in BioRender. Machado, J. (2026) <a target="_blank" href="BioRender.com/vx8zzgy">https://BioRender.com/vx8zzgy</a>.]</p>