Fig. 2

EMID2 inhibits CAF activation and normalizes cancer ECM. A Western blot of total cell lysates and supernatants of HEK293T cells transfected with the indicated plasmids. B Quantification of active-TGFβ, relative to pro-TGFβ. C Western blot of whole muscle tissue injected with LG cells and the indicated vectors. D Quantification of active-TGFβ, relative to pro-TGFβ. Tubulin was used as a loading control. E Primary fibroblasts from COLL-EGFP/αSMA-RFP mice cultured in absence or presence of rEMID2. F Quantification of the percentage of red activated fibroblasts. G Quantification of the area covered by EGFP+/α-SMA⁺ CAFs in muscles co-injected with LG cells and AAV9-CTR or AAV9-EMID2. H Representative images of muscles co-injected with LG cells and AAV9-CTR or AAV9-EMID2. I Representative images of LG tumors in muscles injected with AAV9-CTR and AAV9-EMID2 stained for collagen I. J Representative images of LG tumors in muscles injected with AAV9-CTR and AAV9-EMID2 stained for fibronectin. K. Quantification of collagen I⁺ area as in H. L. Quantification of fibronectin⁺ area as in J. M. Western blots showing collagen I (Col I) content in muscles co-injected with LG cells and AAV9-CTR or AAV9-EMID2. N. Western blots showing fibronectin (Fn) content in muscles co-injected with LG cells and AAV9-CTR or AAV9-EMID2. Tubulin (Tub) was used as loading control. O. Quantification of collagen I as in M. P. Quantification of fibronectin as in N. Scale bar in E 50 μm, in H 1 mm, in I, J 100 μm. In B, D, F, G, K, L, O, P data are shown as mean ± SEM and were analyzed by one-way ANOVA with Student-Newman-Keuls correction. *p < 0.05, **p < 0.01