Fig 2: GLP regulated the expression of cell apoptosis and autophagy. (A) Western blot analysis of the representative proteins; (B) quantification of the expression of C-Caspase 3/Caspase 3, Bcl-2/Bax, NRP-1, P62, LC-3. GAPDH was used as a control. Data are present as mean ± SD (n = 10). ##p < 0.01 as compared to Sham; *p < 0.05, **p < 0.01 as compared to Model.

Fig 3: YY-moxi alone and YY-moxi combined with NAC significantly downregulated NF-κB/p65 levels, increased Bcl-2, and decreased Bax and Caspase-3 levels in MCAO model rats. (A) The levels of Bax, Bcl-2, Caspase-3, and p56 were assessed by IHC assays in the brain tissue of each group. Magnification, 40×; scale bar =50 µm. (B) The positive areas of Bax, Bcl-2, Caspase-3, and p56 were quantitatively analyzed according to the IHC results. (C) Protein expression of Bax, Bcl-2, Caspase-3, and p56 in the injured brain tissue of each group. (D) Western blot analyses of the apoptosis-associated in the MCAO model rats. *, P<0.05, **, P<0.01 vs. the MCAO group; ##, P<0.01 vs. the YY-moxi group. IHC, immunohistochemical; MCAO, middle cerebral artery occlusion model; NAC, N-acetylcysteine; YY-moxi, YaoYi-moxibustion.

Fig 4: Radiation induced cell cycle arrest, autophagy and apoptotic cell death. a Histogram of DNA content in cells reflecting the G0/1, S, and G2/M phases of the cell cycle at 72 h post-irradiation. Stacked column graph shows mean percentage relative (%) ± SEM of triplicates from three independent experiments. b Histogram of autophagosome marker LC3B intensity in indicating the induction of autophagy following irradiation at 72 h post-irradiation. Grey overlay shows the profile at 0 Gy. Column graph shows mean ratio of autophagy induction relative to 0 Gy ± SEM of duplicates from three independent experiments. c Immunoblot of p62, LC3-I and II, and reference protein ß-actin at 72 h post-irradiation. Band intensity was data standardised to ß-actin per sample and then expressed relative to 0 Gy. Raw data is provided in Additional file 4: Fig. S2. Column graph shows mean fluorescent intensity relative to 0 Gy ± SD of from three independent experiments. d Dot plots of apoptotic marker annexin V and viability marker 7-AAD at 72 h post-irradiation. e Dot plots of apoptotic marker caspase 3/7 and viability marker 7-AAD at 72 h post-irradiation. Column and stacked column graphs show mean percentage (%) ± SEM of duplicates from three independent experiments. f Immunoblot of caspase 3 and reference protein ß-actin at 72 h post-irradiation. Band intensity was data standardised to ß-actin per sample and then expressed relative to 0 Gy. Raw data is provided in Additional file 4: Fig. S2. Column graph shows mean fluorescent intensity relative to 0 Gy ± SD of from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 vs 0 Gy determined by two-way ANOVA with Dunnett’s multiple comparison test

Fig 5: Risperidone causes an inhibition of autophagy and a promotion of apoptosis in femur tissues of mice. Mice were treated with 0, 0.3, 0.45, 0.6, 0.75 mg/kg of Risperidone. A representative micro-CT 3D and plan scan images of cross-sectional area of femur tissues. B statistical analysis of BMD in femur tissues of mice assessed by micro-CT. C statistical analysis of BMC in femur tissues of mice. D representative images of HE staining in femur tissues of mice. E Western blot analysis of autophagy-related proteins (LC3 II/I, Beclin1, and p62) expression in femur tissues of mice. The band intensity was assessed. F Western blot analysis of apoptosis-related proteins (cleaved PARP1, cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9). Different numbers represent different concentrations of Risperidone (mg/kg). G co-localization of LC3 II (red), LC3 I (red), Beclin1 (red), and p62 (red) with the osteoblast marker RUNX2 (green) by double-labeled immunofluorescence, wherein the nucleus is labeled by DAPI (blue). *p < 0.05, compared with the treatment of 0 mg/kg of Risperidone. The experiments are repeated 3 times