Figure 2 from Tumor-Secreted ADAMTSL4 Activates Latent TGFβ1 to Drive Cancer Cachexia

No significant differences were observed between groups across the study period. <b>F,</b> Initial and final carcass weight (tumor-free body weight). <b>G,</b> Longitudinal kinetics of relative body weight (BW) change (%) in the days preceding sacrifice. <b>H,</b> Spearman correlation between plasma ADAMTSL4 levels and final relative BW change (<i>n</i> = 18 XY pairs). Relative BW change was calculated from the longitudinal measurements shown in <b>G</b>. <b>I,</b> Relative change in fat mass measured by EchoMRI. <b>J,</b> Fat mass in distinct adipose depots: iWAT, eWAT, and BAT. <b>K,</b> Expression of lipid metabolism-related genes in iWAT assessed by qPCR. <b>L,</b> Relative change in lean mass measured by EchoMRI. <b>M,</b> TA and GC muscle weights. <b>N,</b> Representative images of TA muscle cross-sections stained with WGA Alexa Fluor 488 and DAPI (200× magnification; scale bar, 100 μm).

Representative structural features are highlighted by arrows (white, control; red, ADAMTSL4-overexpressing condition). <b>O,</b> Quantification of muscle fiber CSA. <b>P,</b> Frequency distribution of fiber CSA in TA muscle. <b>Q,</b> Absolute grip strength (forelimbs and hindlimbs). <b>N–P,</b> Complementary readouts derived from the same TA muscle cross-section analysis. <b>R,</b> Grip strength normalized to lean mass (g/g). <b>S,</b> Selected GO terms identified by GSEA in GC muscle from MC38-ADAMTSL4 vs. MC38-GFP tumor-bearing mice (see also Supplementary Data S1 and S2 for full GO term analysis results). <b>T,</b> Validation of marker genes associated with proteasomal catabolic processes (<i>Fbxo32</i> and <i>Trim63</i>). <b>U,</b> Validation of macroautophagy-related genes (<i>Becn1</i>, <i>Map1lc3</i>, and <i>Gabarapl1</i>) by qPCR.

Data are presented as the mean ± SEM. Each dot represents an individual biological replicate. <i>Statistical analysis</i>: two-tailed unpaired <i>t</i> test (<b>B–D</b>, <b>F</b>, <b>I–M</b>, <b>O</b>, <b>Q</b>, <b>R</b>, <b>T</b>, and <b>U</b>), Spearman rank correlation (<b>H</b>), two-way mixed-effects analysis with Šídák multiple-comparisons test (<b>E</b> and <b>G</b>), and weighted Kolmogorov–Smirnov test for GSEA enrichment scores (<b>S</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/q1z1l3n">https://BioRender.com/q1z1l3n</a>.]</p>