Fig. 2

Nexturastat A downregulates SIRPα expression in macrophages. Macrophages were unpolarized (M0, naïve) or polarized to M1-like phenotypes and M2-like phenotype in the presence or absence of Nexturastat A (NextA, 5 µM).A Gene expression analysis of SIRPα in A31A7 macrophages (left panel), BMDMs (middle panel), and THP-1-derived macrophages (right panel) evaluated by qRT-PCR. B Cell surface expression of SIRPα in A31A7 macrophages (left) and BMDMs (right). C Western blots evaluating the SIRPα expression in A31A7 macrophages upon NextA treatment with alpha tubulin as a loading control. D Immunofluorescence microscopy representing changes in SIRPα expression (in green) in BMDMs upon NextA treatment. Nuclei were stained with DAPI (in blue). Scale bars represent 50 μm. E Pearson correlation between HDAC6 and SIRPα expression in skin cutaneous melanoma patients obtained from the TCGA database through GEPIA. F Overall survival of skin cutaneous melanoma patients as it relates to high or low SIRPα expression; data obtained from the TCGA database through GEPIA. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, non-significant