Fig. 1

HDAC6 inhibition modulates macrophage phenotype. Macrophages were unpolarized (M0, naïve) or polarized to M1-like phenotypes and M2-like phenotype in the presence or absence of the HDAC6 inhibitor Nexturastat A (NextA, 5 µM). A Analysis of M1 phenotype markers NOS2 and Cd80 expression of A31A7 macrophages or primary murine BMDMs by qRT-PCR. B Analysis of M1 phenotype cell surface markers H2 and CD80 on A31A7 macrophages or murine BMDMs by flow cytometry. C Analysis of M2 phenotype marker Arg1 expression of A31A7 macrophages or BMDMs by qRT-PCR. D Analysis of M2 phenotype cell surface marker CD206 on A31A7 macrophages or BMDMs by flow cytometry. E Immunofluorescence analysis of M1 marker, iNOS in M1 polarized BMDMs with or without NextA treatment. F Immunofluorescence analysis of M2 marker, Arg1 in M2 polarized BMDMs with or without NextA treatment. Nuclei stained with DAPI are shown in blue, iNOS and Arg1 protein staining are shown in green. G Western blot analysis of M1 (iNOS) and M2 (Arg1) associated markers. Ac-Tubulin indicated inhibition of HDAC6, and Tubulin is loading control. H Gene expression analysis of M1 markers NOS2, CD86, and IL1B in THP-1-derived M1 macrophages and (I) M2 markers MRC1 (CD206) and CD209 in M2 macrophages by qRT-PCR. Scale bars represent 50 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, non-significant