Fig 2: DZX regulates the expression of CHOP, GRP78, caspase-12 and caspase-3 in OGD-treated hippocampal cells. (A) Immunohistochemical staining demonstrated the distribution of GRP78 and CHOP in hippocampal cells exposed to OGD. (B) Western blot analysis was used to detect the expression of GRP78 and CHOP in hippocampal cells exposed to OGD. (C) Protein expression levels of caspase-12 and caspase-3 in hippocampal cells exposed to OGD were detected using western blotting. Data are expressed as the mean ± standard deviation. *P<0.05, as indicated. DZX, diazoxide; CHOP, C/EBP homologous protein; GRP78, 78 kDa glucose-regulated protein; OGD, oxygen-glucose deprivation; 5-HD, 5-hydroxydecanoate.

Fig 3: GM1 reduced neomycin-induced apoptosis in HEI-OC-1 cells. (A) Immunofluorescence images of apoptotic HEI-OC-1 cells with TUNEL and DAPI double staining. (B) Immunofluorescence images of apoptotic HEI-OC-1 cells with cleaved CASPASE-3 and DAPI double staining. (C,D) Quantification of the number of apoptotic cells in (A,B). (E,F) Survival and percentage of HEI-OC-1 cells determined by DAPI/PI staining flow cytometry. (G) Expression of proapoptotic and antiapoptotic genes in HEI-OC-1 cells treated with neomycin and/or GM1 determined by qRT-PCR; the values were normalized to Gapdh as an internal control. The data are expressed as the mean ± SD of triplicate samples. *p < 0.05, **p < 0.01. Scale bars = 20 µm.

Fig 4: Pretreatment with DZX modulates the protein expression of Bcl-2, caspase-12, caspase-3, Bax, CHOP and GRP78 in rats subjected to I/R. Western blot analysis was used to detect the protein expression levels of Bcl-2, caspase-12, caspase-3, Bax, CHOP and GRP78 in hippocampal cells isolated from I/R-treated rats. Rats were treated with DZX with or without 5-HD and subjected to cerebral ischemia for 2 h followed by 12 h of reperfusion. Sham rats were not subjected to cerebral ischemia. DZX, diazoxide; Bcl-2, B-cell lymphoma-2; Bax, Bcl-2-associated X protein; CHOP, C/EBP homologous protein; GRP78, 78 kDa glucose-regulated protein; I/R, ischemia/reperfusion; 5-HD, 5-hydroxydecanoate.

Fig 5: Fe3O4 and MF induce apoptosis of osteoclasts. The amount of pro-apoptotic CASP-3 was elevated in cells treated with Fe3O4 in the presence of MF (A). The Bax:Bcl-2 ratio indicates on the apoptosis activation in cells treated with Fe3O4 and MF (B). Representative photographs from Calcein AM/Propidium Iodide staining (C) and MUSE analysis (D). Results of Muse® Annexin V & Dead Cell assay: number of live (E), early apoptotic (F), late apoptotic/dead (G) and dead cells (H). Results expressed as mean ± SD. Statistical significance indicated as an asterisk (*) when comparing the results between corresponding bars representing MF- and MF+ groups, and a number sign (#) when comparing to CTRL MF-. *,#p<0.05; ###, ***p< 0.001.