Journal: International Dental Journal

Article Title: Tooth Loss Leads to Cognitive Impairment and Mitochondrial Disturbance in Wistar Rats

doi: 10.1016/j.identj.2025.03.027

Figure Lengend Snippet: Autophagy and mitophagy marker changes in the rat hippocampus after tooth loss. (A) Western blot analysis of hippocampal PINK1, P62, parkin, LC3, and GAPDH. (B) Western blot result quantification of hippocampal PINK1, P62, parkin, LC3, and GAPDH (the data are expressed as the mean ± standard deviation [SD], Mann–Whitney U test, * P < .05, ** P < .01, n = 4.)

Article Snippet: The subsequent process included endogenous peroxidase inhibition, blocking, and incubation with primary antibodies against PINK1 (1:2000, Proteintech, 14060-1-AP, China) and parkin (1:600, Proteintech, 23274-1-AP, China).

Techniques: Marker, Western Blot, Standard Deviation, MANN-WHITNEY

Journal: International Dental Journal

Article Title: Tooth Loss Leads to Cognitive Impairment and Mitochondrial Disturbance in Wistar Rats

doi: 10.1016/j.identj.2025.03.027

Figure Lengend Snippet: Immunohistochemical staining and quantitative analysis of PINK1 and parkin in the rat hippocampus. (A) Immunohistochemical staining of PINK1 in the DG, CA3, and CA1 regions of the hippocampus. (B) Quantitative analysis of PINK1 staining intensity in the DG, CA3, and CA1 regions. (C) Immunohistochemical staining of parkin in the DG, CA3, and CA1 regions of the hippocampus. (D) Quantitative analysis of parkin staining intensity in the DG, CA3, and CA1 regions (immunohistochemical staining, × 400 magnification, the mean optical density (MOD) values are expressed as the mean ± standard deviation [SD], Mann–Whitney U test, * P < .05, ** P < .01; n = 3, with analyses conducted on 5 fields per region for each of the 3 animals; scale bar = 50 μm.)

Article Snippet: The subsequent process included endogenous peroxidase inhibition, blocking, and incubation with primary antibodies against PINK1 (1:2000, Proteintech, 14060-1-AP, China) and parkin (1:600, Proteintech, 23274-1-AP, China).

Techniques: Immunohistochemical staining, Staining, Standard Deviation, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: 1α,25(OH) 2 D 3 Promotes the Autophagy of Porcine Ovarian Granulosa Cells as a Protective Mechanism against ROS through the BNIP3/PINK1 Pathway

doi: 10.3390/ijms24054364

Figure Lengend Snippet: 1α,25(OH) 2 D 3 induces mitophagy in PGCs through the ROS-BNIP3-PINK1 signaling pathway. The relative gene expression of BNIP3 ( A , F ) and PINK1 ( B , G ) in PGCs. Western blotting was used to identify the protein level ( C , H ), and the protein contents of BNIP3 ( D , I ) and PINK1 ( E , J ) were evaluated in PGCs. VD 3 : 1α,25(OH) 2 D 3 . NAC: N-acetylcysteine. Different letters on the bar indicate a significant difference. Data are means ± SE of three independent replicates ( p < 0.05).

Article Snippet: Membranes were washed in TBST (150 mM NaCl, 2 mM KCl, 25 mM Tris, 0.05% Tween20, pH 7.4) and incubated with primary antibodies: β-actin (ACTB) (Mouse polyclonal to ACTB, 1:5000 dilution) (Proteintech Group, Inc., Chicago, IL, USA), LC3 (Rabbit polyclonal to LC3I/II, 1:2000 dilution) (Abcam, Cambridge, UK), BNIP3 (Rabbit polyclonal to BNIP3, 1:1000 dilution) (Abcam), PINK1 (Rabbit polyclonal to PINK1, 1:1000 dilution) (Cell Signaling Technology, Danvers, MA, USA), STAR (Rabbit polyclonal to STAR, 1:500 dilution) (Cell Signaling Technology), P62 (Rabbit polyclonal to P62, 1:10,000 dilution) (Servicebio, Wuhan, China) in QuickBlockTM Primary Antibody Dilution Buffer (Beyotime) overnight at 4 °C.

Techniques: Gene Expression, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: 1α,25(OH) 2 D 3 Promotes the Autophagy of Porcine Ovarian Granulosa Cells as a Protective Mechanism against ROS through the BNIP3/PINK1 Pathway

doi: 10.3390/ijms24054364

Figure Lengend Snippet: Porcine-specific Q-PCR primer sequences.

Article Snippet: Membranes were washed in TBST (150 mM NaCl, 2 mM KCl, 25 mM Tris, 0.05% Tween20, pH 7.4) and incubated with primary antibodies: β-actin (ACTB) (Mouse polyclonal to ACTB, 1:5000 dilution) (Proteintech Group, Inc., Chicago, IL, USA), LC3 (Rabbit polyclonal to LC3I/II, 1:2000 dilution) (Abcam, Cambridge, UK), BNIP3 (Rabbit polyclonal to BNIP3, 1:1000 dilution) (Abcam), PINK1 (Rabbit polyclonal to PINK1, 1:1000 dilution) (Cell Signaling Technology, Danvers, MA, USA), STAR (Rabbit polyclonal to STAR, 1:500 dilution) (Cell Signaling Technology), P62 (Rabbit polyclonal to P62, 1:10,000 dilution) (Servicebio, Wuhan, China) in QuickBlockTM Primary Antibody Dilution Buffer (Beyotime) overnight at 4 °C.

Techniques: Sequencing

Journal: Scientific Reports

Article Title: Loss of the mitochondrial kinase PINK1 does not alter platelet function

doi: 10.1038/s41598-018-32716-4

Figure Lengend Snippet: Characterisation of mitochondrial content and function in PINK1 −/− platelets. ( a ) Scanned, full-length agarose gel image showing PINK1 mRNA in WT platelets and absence in KO platelets, with cropped GAPDH mRNA gel as loading control. A full-length gel image for GAPDH is provided in Supplementary Fig. . Samples are from the same experiment and the gels were ran in parallel. ‘No RT’ denotes no reverse transcriptase, negative controls during cDNA synthesis step. ( b ) Cropped immunoblots showing PINK1 expression in 10 µM CCCP (6 hours) treated WT, but not KO platelets by IP (upper panel) with loading control of IP inputs by blotting for PINK1 (lower panel). Uncropped blots for PINK1 are provided in Supplementary Fig. . ( c – e ) Assessment of mitochondrial mass in platelet lysates by citrate synthase activity measurement and detection of mitochondrial markers, VDAC1 and COX IV, by western blotting, with talin as loading control. Cropped blot images with molecular weight markers are shown, with full-length blot at low and high exposure shown in Supplementary Fig. . Samples are from the same experiment and blot. Densitometric analysis of VDAC and COX IV expression levels was expressed as KO:WT ratio, normalised for loading with talin. ( f – i ) Flow cytometry experiments in WT and KO washed platelets (2 × 10 7 /mL) were performed to monitor; ( f ) mPTP opening under basal conditions by calcein quenching with 1 mM cobalt chloride; ( g ) Δψ m with 0.5 µM TMRM; ( h ) reactive oxygen species generation with 5 µM CM-H 2 DCFDA and ( i ) mitochondrial calcium ([Ca 2+ ] M ) levels with 5 µM Rhod-2/AM in response to platelet stimulation for 15 min with CRP (5 µg/mL), thrombin (0.5 U/mL) or combined CRP + thrombin (5 µg/mL + 0.5 U/mL, respectively) in the presence of 1 mM CaCl 2 . Data are mean ± s.e.m (n = 3 for ( c , e ), n = 5 for ( f , g , and i ), n = 7 for ( h )); *p < 0.05, **p < 0.01, ***p < 0.001 vs. basal (or as indicated); (ns) not significant. Median fluorescent intensity (MFI) values are reported for ( g – i ).

Article Snippet: For western blotting, the antibodies were as follows: PINK1 (#BC100–494, 1:1000 dilution, NOVUS Biologicals, Abingdon, U.K.), PINK1 (#sc-33796, 1:500 dilution, Santa Cruz), PINK1 (#S774C and #S086D, both 1:500 dilution, Dundee MRC PPU), PINK1 (#6946, 1:1000 dilution), voltage dependent anion channel (VDAC, #4866, 1:1000 dilution) and cytochrome oxidase (COX) IV (#4850, 1:1000 dilution) antibodies were from Cell Signaling Technology (New England Biolabs, Hitchin, UK) and talin antibody (#sc-7534, 1:500 dilution) was from Santa Cruz.

Techniques: Agarose Gel Electrophoresis, Control, Reverse Transcription, cDNA Synthesis, Western Blot, Expressing, Activity Assay, Molecular Weight, Flow Cytometry