Figure EV3. Therapeutic ADAM12 vaccination targets ADAM12⁺ CAFs and inhibits cell proliferation in subcutaneous PDAC tumors.

(A) Representative immunofluorescence staining images (left) and quantitative analysis (right) of ADAM12⁺/α-SMA⁺ (left, left), ADAM12⁺/PDGFR-β⁺ (left, middle), and ADAM12⁺/CK19⁺ (left, right) co-expression on PDAC tumors from control-vaccinated mice (v-CTRL) and ADAM12-vaccinated mice (v-A12). Data were represented as the number of cells per high power field. Representative double positive cells were indicated by white arrows respectively. (n = 7–8 mice in v-CTRL, n = 8 mice in v-A12. Data were presented as mean ± SEM. Statistical test: multiple t-test. Scale bar 50 μm). (B) Representative immunofluorescence images (left) and quantitative analysis (right) of proliferating (Ki67⁺) ADAM12⁺ cells on PDAC tumors from v-CTRL mice and v-A12 mice. (n = 7 mice in v-CTRL, n = 6 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t-test. Scale bar 50 μm). (C) Representative immunofluorescence staining images (left) and quantitative analysis (right) of proliferating (Ki67⁺) tumor (CK19⁺) cells on PDAC tumors from v-CTRL mice (n = 7) and v-A12 mice (n = 6). (n = 7 mice in v-CTRL, n = 6 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t-test. Scale bar 50 μm). (D) Scheme of therapeutic vaccination with gemcitabine treatment on subcutaneous KP2 PDAC tumor-bearing mice. (E) Representative immunofluorescence images (left) and quantitative analysis (right) of apoptotic cells (cleaved caspase 3⁺) on PDAC tumors from v-CTRL mice and v-A12 mice. (n = 5 mice in v-CTRL, n = 6 mice in v-A12. Data were presented as mean ± SEM. Statistical test: unpaired two-tailed Student’s t-test. Scale bar 50 μm).