Fig 2: Caspase-2 plays a dominant role in EV71-induced INF2 cleavage. (A) RD cells were transfected with control siRNA or siRNA against caspase-2, -3, and -8, and 36 h after transfection, the cells were mock-infected (EV71-) or infected with EV71 (MOI = 10) for an additional 12 h. Then, the cells were harvested, and a Western blot was performed to detect INF2, the corresponding caspases, actin, and VP1. (B) RD cells were mock-infected (–) or infected with EV71 (MOI = 10) for 12 h, 100 µM Z-VAD-FMK (Z-VAD, pan-caspase inhibitor), Z-DEVD-FMK (Z-DEVD, caspase-3 inhibitor), Z-IETD-FMK (Z-IETD, caspase-8 inhibitor) (the left panel), and 30 or 60 µM Z-VDVAD-FMK (Z-VDVAD, caspase-2 inhibitor) were added to the culture media 6 h post-infection. Then, the cells were harvested and lysed for Western blot to detect INF2, PARP, Actin, and EV71 VP1. PARP was used as an indicator of caspase activation and the effect of caspase inhibitors. Arrows indicate the cleavage fragments (CF). (C) RD cells, caspase-2 knockout (KO) cells, caspase-2, -3 double KO cells, and caspase-3 KO were infected with EV71 (MOI = 10) for 12 h; then the cells were harvested, and Western blot was conducted to detect INF2, caspase-2, caspase-3, actin, and EV71 VP1.

Fig 3: r-PGRN reduced neuronal apoptosis and improved synaptic development, while PI3K inhibitor wortmannin reversed this effect in VPA-induced rats.Representative blots (A) and quantification (B) of Caspase-3, BCL-2, and Bax in the four groups. C, D Representative images (bar = 50 µm) and quantification of TUNEL staining. Representative blots (E) and quantification (F) of synaptic markers such as PSD95 and SYP. G, H Immunofluorescence staining (bar = 25 µm) exhibited consistent changes in PSD95 and SYP protein expressions. Representative images of Purkinje cell dendrites (I) that were used for quantification of spines (bar = 10 µm) and quantification (J) of the number of spines. Data are expressed as mean ± SEM. One-way ANOVA followed by Tukey’s post hoc (B(Caspase-3, Bax), D, F(SYP), H); post hoc Kruskal–Wallis test for multiple comparisons (B(Bcl2), F(PSD95), J). (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, sample sizes (n): n = 5/group).

Fig 4: TMPO-AS1 regulates BC growth in vivo via E2F1. (A) Representative images of the tumor xenografts 25 days after the subcutaneous injection of RT4 cells transfected with empty vector, sh-TMPO-AS1, and sh-TMPO-AS1 along with an E2F1-overexpression construct into the flanks of nude mice. (B) The tumor volumes were measured every 3 days in the three groups. Three independent experiments were performed. (C) The expression of E2F1, Ki-67, and caspase-3 in xenografts was evaluated using immunohistochemistry. Representative images are shown. (D) A schematic diagram illustrating the role and mechanism of TMPO-AS1 in BC tumorigenesis and progression is illustrated. *p < 0.05.

Fig 5: SCFA supplementation mitigates neuroinflammation and promotes neuronal survival in the hippocampus post BCCAO. A, B Representative immunofluorescence images showing Iba-1+ cells in the hippocampus at day 14 post BCCAO (A) and the quantification analysis (B) (n = 6/group). Scale bar, 50 µm. C, D mRNA levels of inflammatory cytokines (IL-4, IL-10, TNF-a, IL-1ß, and IL-6) in the hippocampus at day 14 post BCCAO (n = 5/group). E Protein levels of CD16 and NF-?B and their quantification in the hippocampus at day 14 post BCCAO (band intensity normalized to ß-actin) (n = 6/group). F, G Representative immunofluorescence images (F) and mean fluorescence intensity (G) of hippocampal neurons in CA1, CA2, CA3, and DG in the different groups. (n = 6/group). Scale bar, 50 µm. H–K Western blot analysis for protein levels of caspase 3, cleaved caspase 3, Bcl2, Bax, Bcl-XL, Erk, and pErk and their quantification (band intensity normalized to ß-actin or COXIV) in the hippocampus (n = 6/group). The data represent the mean ± SEM. p < 0.05 was set as the threshold for significance. * p < 0.05, ** p < 0.01, *** p < 0.001 compared to S+NaCl group. &&p < 0.01 compared to the S+SCFA group. +p < 0.05 compared to the B+NaCl 14d group. #p < 0.05, ##p < 0.01 compared to the B+NaCl 37d group