Figure 1. The design of chemo-immuno therapy.
(A) Masson’s trichrome staining illustrating desmoplastic type of melanoma with collagen-rich TME (blue-stained collagen highlighted with black arrows), compared with relatively non-desmoplastic samples from patients and mouse tumor specimens. Scale bar indicates 300 μm. (B) Flow cytometry analysis showing the major cell populations within TME. (Tumor cells: MART1+; TAFs: FAP+). (C) A total of 25 candidate drugs were screened by MTT assay. MIT and CEL were selected among lowest IC50. n = 5. (D) IC50 and CI of MIT, CEL, and optimized ratios of MIT+CEL on both BPD6 cells (desmoplastic melanoma cells) and 3T3-T cells (TGF-β activated fibroblast, mimicking TAFs in vivo). 24 h incubation. n = 5–8. (E) ICD induced by MIT and MIT+CEL (combination ratio of 5:1) on tumor cells. For MIT: 16 μM, for MIT+CEL: 4.5 μM of MIT and 0.9 μM of CEL. Fluorescence imaging detecting ICD markers: CRT and HMGB1. Cell nuclei were stained with Hoechst 33432. Scale bar indicates 10 μm. n = 3.