Journal: Scientific reports

Article Title: Attenuated PINK1 autophosphorylation play neuroprotective and anti-seizure roles in neonatal hypoxia.

doi: 10.1038/s41598-025-99915-8

Figure Lengend Snippet: Fig. 3. Inhibition of TOM7 expression reduced the levels of phosphorylated PINK1/Parkin/Ub during hypoxia-induced seizures. (A,B) TOM7 (green) immunofluorescence intensity (3 day; n = 4/group). Bar = 50 μm. (C-H) Changed levels of PINK1/P-PINK1 and Parkin/P-Parkin due to TOM7 intervention (n = 4/ group). (I-J) P-Ub (red) immunofluorescence intensity in the EC and DG (3 day; n = 4/group). DAPI, blue. Bar = 50 μm. Mean ± SEM are shown. Compared with controls, *P < 0.05, **P < 0.01, and ***P < 0.001 (one-way ANOVA). ###P < 0.001 compared with each other (one-way ANOVA with Dunnett’s T3 post-hoc test). C cortex, DG dentate gyrus, EC entorhinal cortex, H hippocampus, Hy Hypoxia, P-Parkin phosphorylated Parkin, P-PINK1 phosphorylated PTEN-induced kinase 1, P-Ub phosphorylated ubiquitin, TOM7 translocase outer mitochondrial membrane 7.

Article Snippet: Brain sections were washed with 0.01 M phosphate buffered saline (PBS) and incubated with primary antibody, including mouse monoclonal antibody against TOM7 (1:200; 15071-1-AP; Proteintech, USA), LC3B (1:200; ab48394; Abcam), TOMM20 (1:200; ab56783; Abcam), and rabbit monoclonal antibody against Ser-65-phospho-Ub (1:200; 70973, CST) at 4°C overnight respectively.

Techniques: Inhibition, Expressing, Immunofluorescence, Ubiquitin Proteomics, Membrane

Journal: Scientific reports

Article Title: Attenuated PINK1 autophosphorylation play neuroprotective and anti-seizure roles in neonatal hypoxia.

doi: 10.1038/s41598-025-99915-8

Figure Lengend Snippet: Fig. 4. Inhibition of TOM7 expression attenuated mitophagy, apoptosis, mitochondrial oxidative stress and neuronal damage. (A-C) Inhibition of TOM7 expression significantly reduced the fluorescence intensity of LC3B (red) and TOMM20 (green) in the EC and DG (3 day; n = 4/group). DAPI, blue. Bar = 30 μm. (D-H) The changed levels of P62, active caspase-3 and caspase-3 in the cortex and hippocampus caused by TOM7 intervention (n = 4/group). (I-L) Mito-SOX levels in the cortex and hippocampus detected by fluorescence microplate reader and flow cytometry (n = 4/group). (M,N) Neuronal damage in the DG, CA3 and EC (3 day; n = 4/group). Bar = 50 μm. Mean ± SEM are shown. Compared with controls, *P < 0.05, **P < 0.01, and ***P < 0.001 (one-way ANOVA); and #P < 0.05, ##P < 0.01, compared with each other (one-way ANOVA with Dunnett’s T3 post-hoc test). C cortex, CA3 cornu ammonis area 3, DG dentate gyrus, EC entorhinal cortex, H hippocampus, Hy Hypoxia, LC3B microtubule-associated protein light chain 3B, TOMM20 translocase of outer mitochondrial membrane 20, TOM7 translocase outer mitochondrial membrane 7.

Article Snippet: Brain sections were washed with 0.01 M phosphate buffered saline (PBS) and incubated with primary antibody, including mouse monoclonal antibody against TOM7 (1:200; 15071-1-AP; Proteintech, USA), LC3B (1:200; ab48394; Abcam), TOMM20 (1:200; ab56783; Abcam), and rabbit monoclonal antibody against Ser-65-phospho-Ub (1:200; 70973, CST) at 4°C overnight respectively.

Techniques: Inhibition, Expressing, Fluorescence, Flow Cytometry, Membrane

Journal: Scientific reports

Article Title: Attenuated PINK1 autophosphorylation play neuroprotective and anti-seizure roles in neonatal hypoxia.

doi: 10.1038/s41598-025-99915-8

Figure Lengend Snippet: Fig. 5. Inhibition of TOM7 expression attenuated seizures, learning and memory defects. (A,B) Number and cumulative duration of behavorial seizures. (C,D) Seizure number and cumulative seizure duration of EEGs (nonparametric Mann–Whitney U test). (E-G) Representative EEGs and PSD analyses of each group. (H) Target quadrant time (%). (I) Opposite quadrant time (%). (J) Target zone frequency. (K) Latency to platform (two-way ANOVA with Dunnett’s T3 post-hoc test). Control group, n = 11; Hypoxia negative vector group, n = 12; TOM7 intervention group, n = 14. Data are presented as mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001, Compared with controls (one-way ANOVA); and #P < 0.05, ###P < 0.001; compared with each other (one-way ANOVA with Dunnett’s T3 post-hoc test). EEGs electroencephalograms, PSD power spectrum density, TOM7 translocase outer mitochondrial membrane 7.

Article Snippet: Brain sections were washed with 0.01 M phosphate buffered saline (PBS) and incubated with primary antibody, including mouse monoclonal antibody against TOM7 (1:200; 15071-1-AP; Proteintech, USA), LC3B (1:200; ab48394; Abcam), TOMM20 (1:200; ab56783; Abcam), and rabbit monoclonal antibody against Ser-65-phospho-Ub (1:200; 70973, CST) at 4°C overnight respectively.

Techniques: Inhibition, Expressing, MANN-WHITNEY, Control, Plasmid Preparation, Membrane