Fig 2: Inhibition of HDAC7 rescues the function of miR-143 deficiency in mice.qRT-PCR analysis (a) of the relative expression levels of HDAC7 and western blot analysis (b). qRT-PCR analysis (c) of the relative levels of HDAC7 and Western blot analysis (d) of HDAC7 and GADPH in BMSCs derived from WT, miR-143 knockout mice transfected with HDAC7-siRNA, and miR-143-knockout male mice (n = 8 per group). Microcomputed tomography images (e) of trabecular bone microarchitecture in femora and quantitative microcomputed tomography analysis (f) of trabecular bone microarchitecture from miR-143-knockout mice injected with cholesterol-conjugated control RNA or HDAC7-siRNA in the tail vein (n = 8 per group). Scale bar: 100 μm. g, h Representative images of calcein double labeling of trabecular bone (g) with quantification of the BFR per bone surface (BFR/BS) (h left) and the mineral apposition rate (MAR) (h right) (n = 8 per group). Scale bar: 100 μm. i Representative images of CD31 (green), Emcn (red) immunostaining from miR-143-knockout mice injected with cholesterol-conjugated control RNA or HDAC7-siRNA in the tail vein. Scale bar: 100 μm. j FACS analysis dot plot of CD31hiEmcnhi endothelial cells from miR-143-knockout mice injected with cholesterol-conjugated control RNA or HDAC7-siRNA in the tail vein. Data are reported as the mean ± SD. *p < 0.05, **p < 0.01. Data are representative of three independent experiments.

Fig 3: Epigenetic modifiers increase γδ T cell treatment efficacy. (A) Dose–response curves of bicellular A375‐fibroblast spheroids, A2058‐fibroblast spheroids, patient derived melanoma spheroids and melanoma patient‐derived organoids (MPDOs) after treatment with Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat, Vorinostat and DMSO controls for 3 days. Values represent mean ± SEM, n = 4. (B) Expression of MICA/B, FasL, PD‐L1 and PD‐L2 in cells from A2058‐fibroblast spheroids was measured by FACS after Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat and Vorinostat treatment (5 μM) for 48 h. *p < 0.05. **p < 0.01. (C) Western blot analysis of HDAC6, HDAC7, COX2, phospho‐STAT3, total STAT3, phosphor‐ERK, total ERK, phospho‐JNK, total JNK and GAPDH proteins in A2058‐fibroblast spheroids after Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat and Vorinostat treatment (5 μM) for 48 h (left panel). Densitometry quantification of the HDAC7, HDAC6 and COX2 (right panel). **p < 0.01. (D) NKG2A, PD‐1, NKG2D, CXCR3. CD107a and intracellular IFN‐γ in γδ T cells was measured by FACS after Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat and Vorinostat treatment (5 μM) for 48 h. *p < 0.05. **p < 0.01. (E) Cytotoxic function of γδ T cells against A2058‐fibro spheroids and MPDOs combined with 1 μM of Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat, Vorinostat and DMSO controls. n = 4. *p < 0.05. **p < 0.01

Fig 4: HDAC7 promotes ovarian cancer malignant progression by regulating AKT/mTOR signal pathway. (A) The levels of AKT/mTOR signal pathway were measured by Western blot, after treated with MK‐2206 2HCl for 24 h; (B–D) The cell proliferation (B), Colony formation assay (C) and Transwell migration assay (D) of ES2 and OVCAR‐3 cell lines treated with MK‐2206 2HCl; The cell experiment was repeated 3 times. The data were expressed as mean ± standard deviation and analysed by the t test (*p < 0.05; **p < 0.01; ***p < 0.001).

Fig 5: Ectopic expression of HDAC7‐mut eliminates the effects of HDAC7 knockout in ovarian cancer cells. (A) The levels of AKT/mTOR signal pathway and HDAC7 were measured by Western blot in HDAC7 knockout or HDAC7‐mut and control groups. Cells with HDAC7‐mut show rescue the level of p‐AKT(S473) and p‐mTOR(S2448), compared to HDAC7 knockout groups; (B–D) The cell proliferation (B), Colony formation assay (C) and Transwell migration assay (D) of ES2 and OVCAR‐3 cell lines which transfected HDAC7 knockout or HDAC7‐mut Lentiviral; The cell experiment was repeated 3 times. The data were expressed as mean ± standard deviation and analysed by the t test (*p < 0.05; **p < 0.01; ***p < 0.001).