Fig. 2

CP1 induces immunogenic cell death while increasing pro-inflammatory cytokines and chemokines and decreasing VEGF. ICD was assessed in vitro from co-culture of a Myc-CaP or b LNCaP cells with untreated (Unt.), mitoxantrone (Mx), heat killed (HK) CP1, or live CP1 via HMGB1 (ELISA), ATP (luminescence assay), and calreticulin (flow cytometry, with representative histogram (Unt = black, Mx = gray, CP1 HK = dark red, CP1 live = red)), performed in biological triplicates, technical duplicates, statistics compared to Unt. ICD was assessed in vivo by c HMGB1 or d calreticulin IF of prostate tumor tissue 9 days after intra-urethral CP1 administration, with representative images (each calreticulin image representative of a different tumor with white arrows indicating foci of cell surface staining, green = HMGB1 or calreticulin, scale bar, 50 μm). Mice n = 4/group, HMGB1 quantified with quadruplicate FOVs/tumor. e Multiplex cytokine and chemokine array from Myc-CaP supernatant, performed in biological triplicates, technical duplicates. Data represented as mean ± S.E.M. or log₂ fold change with and without CP1 exposure. Statistical significance was determined by two-tailed Student’s t-test (a, b, each group compared to Unt.). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001