Fig. 3

FAK inhibition propels ICD biomarker exposure in the presence of doxorubicin on the cell lines of different cancer types. A The most significantly altered signaling pathways upon combination treatment with IN10018 and doxorubicin. SK-OV-3 cells were treated with DMSO, 3 μM IN10018, 300 nM doxorubicin, and a combination of 3 μM IN10018 and 300 nM doxorubicin for 24 h. RNA sequencing and wiki pathway analysis were performed for the combination treatment group compared to doxorubicin monotherapy group here. The items marked in blue indicate ICD-related signaling and those marked in red indicate FAK downstream. B The combination of IN10018 and doxorubicin increased the expression of genes which can boost the cancer cell-killing effects CD8 positive T cells. C The apoptosis of SK-OV-3 cells induced by the treatment with DMSO, 3 μM IN10018, 300 nM doxorubicin, and the 2-drug combination for 48 h. (n = 3 per group). D Calreticulin releasing in the cell culture supernatant of the treated SK-OV-3 cells from (C). E–F Calreticulin and HMGB1 staining for treated SK-OV-3 cells from (C) (n = 3 per group). G ATP-releasing percentage of treated SK-OV-3 cells treated with indicated drugs in (C) for 24 h (n = 3 per group). H-P The annexin V/PI, calreticulin, and GRP94 staining in the mouse breast cancer cell line 4T1 (H-J), moues colorectal cancer cell line CT26 (K-M), and mouse ovarian cancer cell line ID8 (N-P) treated with different therapeutics for 48 h. The cells were stained with each antibody and analyzed using flow cytometry (n = 3 per point). Data represent mean ± SEM. Statistical analysis was done using the unpaired student's T-test. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001