Fig 2: Effect of miR-29a-3p on the proliferation and apoptosis levels of glioma cells under normoxic and hypoxic conditions. (A) The expression level of miR-29a-3p by reverse transcription-quantitative PCR, ***P<0.001 vs. miR-NC under hypoxia. (B) The effect of miR-29a-3p on the proliferation level of glioma cells U251, ####P<0.0001 vs. miR-NC under normoxia; ***P<0.001, ****P<0.0001 vs. miR-NC under hypoxia. (C) Effect of miR-29a-3p on apoptotic levels in glioma cells U251, *P<0.05 vs. miR-NC under hypoxia, ***P<0.001 vs. miR-NC under normoxia. (D) Effect of miR-29a-3p on the expression levels of Bax, Bcl-2 and HIF-1α. *P<0.05, **P<0.01. All data were expressed as the mean ± standard deviation (n=3). miR, microRNA; NC, negative control; HIF-1α, hypoxia-inducible factor 1.

Fig 3: Electrotaxis of BEAS-2B cells and the antiapoptotic potential of EFs. (A) Time-lapse photographs of BEAS-2B cells in the control group and the experimental group at an EF of 200 mV/mm, with red lines and yellow arrowheads as cell migration routes and their directions; (B) Trajectories of the migration of BEAS-2B cells with EFs at 0 and 200 mV/mm, with black lines as cells migrating toward the cathode and red lines vice versa (or left and right in the control, no EF); (C) Velocity and directionality of cellular migration in EFs. (D) Immunofluorescence staining of BEAS-2B cells with EFs of 0 and 200 mV/mm; (E) WBA showing the difference in the phosphorylation of AKT after application with EFs in BEAS-2B cells; (F) WBA showing the difference in apoptosis-related molecules after application with EFs in BEAS-2B cells.∗p < 0.001 compared with the control.BEAS-2B cells: human bronchial epithelial cells transformed with Ad12-SV40 2B; WBA: western blot analysis; EF: electric field; Ctrl: control, 0 mV/mm; DAPI: 4′,6-diamidino-2-phenylindole; P-AKT: phosphorylated AKT; P-MCL-1: phosphorylated myeloid leukemia-1; Bcl-xl: B-cell lymphoma-extra large; Bax: Bcl-2-associated X protein; Bim: Bcl-2-like protein 11; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.

Fig 4: DHPITO activates apoptosis in both HCT116 and HCT8 cell lines. (A) Flow cytometric analysis using annexin V/PI staining indicates that cell apoptosis was induced by DHPITO in HCT116 and HCT8 cells. (B) Protein levels of cleaved PARP, cytochrome c, BCL2, BAX, and cleaved-caspase 8 detected with Western blotting following treatment with the indicated concentrations of DHPITO for 24 h. GAPDH was used as a loading control. Ns, no significance. * p < 0.05; ** p < 0.01; *** and **** p < 0.001 versus vehicle. PI, propidium iodide; PARP, poly ADP-ribose polymerase; BCL2, B-cell lymphoma 2; BAX, BCL2 -associated X protein.

Fig 5: SENP3 deletion alleviated the neurotoxic effects of pro-inflammatory microglia in cocultured neurons after OGD/R(A) Primary cultured microglia were transduced with Ad-SENP3 or Ad-sh.SENP3, then co-cultured with primary neurons via transwell system, following by OGD/R treatment. Schematic representation as above.(B and C) The apoptotic neurons were determined by TUNEL staining (B) and quantification analysis of TUNEL+ neurons (C). Scale bar: 20 μm.(D) The cytotoxicity was examined by LDH release assay.(E) The neuronal viability was determined by CCK-8 assay.(F–K) (F) Immunoblot assays showed the protein expression of representative anti-apoptosis or pro-apoptosis molecules in co-cultured neurons. (G-K) Quantitative analysis of the Bcl-xl (G), Bax (H), cleaved PARP (I), cleaved caspase-9 (J), and cleaved caspase-3 (K) protein expression in (F). Data are presented as the means ± SEM. from at least three dependent experiments and analyzed by two-way ANOVA followed by Tukey’s post hoc test. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001.