Fig 2: The schematic depicts the functional mechanisms of WBM in hypoxia-caused damage in microglial BV-2 cells. (a) Hypoxia increased ferroptosis and apoptosis via regulations of GPX4/SLC7A11 and cleaved-caspase3/Bax/Bcl-2, respectively. Moreover, hypoxia also induced ER stress via the regulation of CHOP/Bip. (b) WBM reversed the hypoxia-triggered ferroptosis and apoptosis via regulations of GPX4/SLC7A11 and cleaved-caspase3/Bax/Bcl-2, respectively. Conversely, WBM also alleviated hypoxia-induced ER stress via the regulation of CHOP/Bip. This figure was created with https://www.BioRender.com.

Fig 3: circPHKA2 inhibited apoptosis, ER stress, and oxidative stress of HBMEC after OGD. OGD-induced HBMEC were pretransfected with oe-circPHKA2 or NC, compared with control HBMEC transfected with NC. (a) FCM identified percentage of apoptotic cells. (b, c) Western blotting detected relative protein expression of Bcl-2, Bax, GRP78, CHOP, caspase-12, and ß-actin. (d–f) Assay kits of ROS, MDA, and SOD examined relative ROS level, MDA level, and SOD activity, respectively. (g) Western blotting detected relative SOD2 protein expression, normalized to ß-actin. *P < 0.05.

Fig 4: MHO7 induced ER stress and cell cycle arrest in TNBC cells. (A) The top KEGG pathway enrichment was analyzed by RNA-seq. The red highlighted upregulated pathways, and the blue highlighted downregulated pathways. (B) The heat map of DEGs related to ER stress. (C) ER stress-regulated genes in MDA-MB-231 cells were detected by qRT-PCR under MHO7 treatment. (D) The expression of BiP/p-PERK/p-eIF2a/ATF4/CHOP were measured by western blot under MHO7 treatment in MDA-MB-231 cells. (E) Bcl-2 was detected by qRT-PCR when si-CHOP or MHO7 (3 µM) treatment. (F) Bax was detected by qRT-PCR when si-CHOP or MHO7 (3 µM) treatment. (G) Representative apoptosis rate was measured by flow cytometry under si-CHOP or MHO7 (3 µM) treatment. *P <0.05; **P < 0.01; ***P <0.001; and ****P < 0.0001; ns=no significant.

Fig 5: circPHKA2 inhibited apoptosis, ER stress, and oxidative stress of HBMEC after OGD by modulating SOD2. OGD-induced HBMEC were pretransfected with oe-circPHKA2, NC, or oe-circPHKA2 combined with commercial miR-574-5p or miR-NC. (a) FCM identified percentage of apoptotic cells. (b, c) Western blotting detected relative protein expression of Bcl-2, Bax, GRP78, CHOP, caspase-12, and ß-actin. (d–f) Assay kits of ROS, MDA, and SOD examined relative ROS level, MDA level, and SOD activity, respectively. *P < 0.05.