Journal: Frontiers in Endocrinology

Article Title: Transcriptome Analysis Reveal Candidate Genes and Pathways Responses to Lactate Dehydrogenase Inhibition (Oxamate) in Hyperglycemic Human Renal Proximal Epithelial Tubular Cells

doi: 10.3389/fendo.2022.785605

Figure Lengend Snippet: The protein expression levels of peroxisome proliferator-activated receptor-g coactivator 1α (PGC1α), caspase 3 (CASP3), B-cell lymphoma 2 apoptosis regulator (BCL2), BCL2 associated X apoptosis regulator (BAX), beclin1 (BECN1) and microtubule-associated proteins 1A/1B light chain 3 (MAP1LC3) in HK-2 cells treated with low D-glucose (LG) with different concentrations of oxamate (LGOXA-0mM, LGOXA-20mM, LGOXA-40mM and LGOXA-80mM) and high D-glucose (HG) with different concentrations of oxamate (HGOXA-0mM, HGOXA-20mM, HGOXA-40mM and HGOXA-80mM) for 24 hours. (A) Western blot gels. (B) Relative grey values. * indicates P -value < 0.05 after student’s t -test.

Article Snippet: The primary antibodies [Anti-PGC1α (ab54481), Anti-CASP3 (9665), Anti-BCL2 (ab59348), Anti-BAX (GB11690), Anti-BECN1 (bs-1353R), Anti-MAP1LC3 (14600-1-ap) and Anti-β-actin (GB12001)] and all secondary antibodies were purchased from abcom (Cambridge, MA, USA), Proteintech (Wuhan, China), BIOSS (Boston, Massachusetts, USA) and Servicebio (Wuhan, China), respectively.

Techniques: Expressing, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: DPSCs modulate synovial macrophage polarization and efferocytosis via PINK1/Parkin-dependent mitophagy

doi: 10.1186/s13287-025-04468-2

Figure Lengend Snippet: DPSCs promote autophagy and M2 polarization in macrophages. (a) Immunohistochemical analysis of autophagy-associated proteins P62, Beclin1 and LC3 in synovial macrophages of various treatment groups. Scale bar: 10 μm. ( b - d ) Quantifying the histomorphometric analysis of the expression of proteins P62, Beclin1 and LC3. ( e - f ) Immunofluorescence staining of CD86 and CD206 in RAW264.7 macrophages. Scale bar: 10 μm. Data are presented as the mean ± SD. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** P < 0.0001

Article Snippet: Antibodies used for IHC/IF staining were as follows: P62 (HUABio, R1309-8, China), Beclin1 (ABclone, A7353, China), LC3B (Novusbio, NB100-2220, USA), CD206 (Proteintech, 18704-1-AP, China), CD86 (Proteintech, 13395-1-AP, China), Col2 (Abcam, ab39012, UK), MMP13 (Abcam, ab39012, UK).

Techniques: Immunohistochemical staining, Expressing, Immunofluorescence, Staining

Journal: Stem Cell Research & Therapy

Article Title: DPSCs modulate synovial macrophage polarization and efferocytosis via PINK1/Parkin-dependent mitophagy

doi: 10.1186/s13287-025-04468-2

Figure Lengend Snippet: DPSCs enhance macrophage mitochondrial autophagy in vitro . ( a ) The visualization of the ultrastructure of macrophages via transmission electron microscope (TEM). In the images, membrane-like vesicles in macrophages were observed. The distribution of autophagosomes (green circle) in macrophages was visualized using the TEM. Scale bar: 1 μm. Amplification region: 500 nm. ( b , f ) Autophagocytic flux in macrophages was analyzed by mRFP-GFP-LC3 (yellow puncta). Scale bar: 10 μm. ( c - d ) Western blot showing P62, Beclin1 and LC3-II/I expression in macrophages after various treatments. ( e , g ) Western blot showing the expression of PINK1 and Parkin in macrophages after various treatments. Data are presented as the mean ± SD. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** P < 0.0001

Article Snippet: Antibodies used for IHC/IF staining were as follows: P62 (HUABio, R1309-8, China), Beclin1 (ABclone, A7353, China), LC3B (Novusbio, NB100-2220, USA), CD206 (Proteintech, 18704-1-AP, China), CD86 (Proteintech, 13395-1-AP, China), Col2 (Abcam, ab39012, UK), MMP13 (Abcam, ab39012, UK).

Techniques: In Vitro, Transmission Assay, Microscopy, Membrane, Amplification, Western Blot, Expressing

Journal: bioRxiv

Article Title: A mitochondrial SCF-FBXL4 ubiquitin E3 ligase complex restrains excessive mitophagy to prevent mitochondrial disease

doi: 10.1101/2022.11.11.516094

Figure Lengend Snippet: (A) Volcano plot of the screen result of mitophagy regulators in FBXL4-KO HeLa cells. Mitochondrial outer-membrane protein MTCH2 and mitophagy receptors BNIP3 and NIX are highlighted in red. (B) Immunoblot analysis of the indicated HeLa cells. WT and FBXL4-KO HeLa cells were rescued with vector or FBXL4-FLAG. (C) qPCR analysis of BNIP3 and NIX in the indicated HeLa cells. (D) Immunoblot analysis of the indicated HeLa cells. B: BNIP3; N: NIX. Two clones were shown for each double and triple knockout cells. (E and F) Representative live cell imaging (E) and quantitative analysis (F) of mitophagy levels in the indicated HeLa cells. n: number of cells analyzed (left); number of imaging areas analyzed (right). (G) Representative FACS analysis of mitophagy levels in the indicated HeLa cells. (H) FACS-based quantitative analysis of mitophagy levels in the indicated HeLa cells. Data are mean + SD from three biological replicates (C, H). Statistics: two-tailed unpaired Student’s t-test (C, F and H); ** P < 0.01; *** P < 0.001.

Article Snippet: BNIP3 (Abcam, ab109362; Abcam, ab10433; Cell signaling, 3769S), NIX (Cell signaling, 12396S), FUNDC1 (From Quan Chen lab), FLAG (Sigma-Aldrich, F1804), HA (Sigma-Aldrich, H6533), ACTIN (Sigma-Aldrich, A2066), TOM70 (Proteintech, 14528-1-AP), TOM20 (ABclonal, A19403), SMAC (Cell signaling, 2954S), MITOFILIN (Proteintech, 10179-1-AP), HSP60 (Cell signaling, 4870S), TIMM23(Proteintech, 11123-1-AP), SKP1 (Cell signaling, 2156S), CUL1 (Proteintech, 12895-1-AP), Total OXPHOS Cocktail (Abcam, ab110411), Cytochrome C (BD Biosciences, 556433), mCherry (Easybio, BE2026), MTCH2 (Proteintech, 16888-1-AP), BECLIN1 (Cell signaling, 3495T), FIP200 (ABclonal, A14685), TFAM (Abcam, ab131607), LONP1 (Proteintech, 15440-1-AP), VDAC (Cell signaling, 4866S), Donkey polyclonal anti-Rabbit IgG (H+L),HRP-conjugated (Jackson ImmunoResearch, 711-035-152), Donkey polyclonal anti-Mouse IgG (H+L), HRP-conjugated (Jackson ImmunoResearch, 711-035-151), Goat polyclonal anti-Mouse IgG, Fcγ fragment specific, HRP-conjugated (Jackson ImmunoResearch, 115-035-008), Goat polyclonal anti-Mouse IgG, light chain specific, HRP-conjugated (Jackson ImmunoResearch, 115-035-174).

Techniques: Membrane, Western Blot, Plasmid Preparation, Clone Assay, Triple Knockout, Live Cell Imaging, Imaging, Two Tailed Test

Journal: bioRxiv

Article Title: A mitochondrial SCF-FBXL4 ubiquitin E3 ligase complex restrains excessive mitophagy to prevent mitochondrial disease

doi: 10.1101/2022.11.11.516094

Figure Lengend Snippet: (A and B) Representative FACS analysis (A) and quantitative analysis (B) of mitophagy levels in the indicated HeLa cells. Two independent sgRNAs for MTCH2 were used. Data are mean + SD from three biological replicates. Statistics: two-tailed unpaired Student’s t-test; *** P < 0.001. (C) Immunoblot analysis of the indicated HeLa cells. *: non-specific bands.

Article Snippet: BNIP3 (Abcam, ab109362; Abcam, ab10433; Cell signaling, 3769S), NIX (Cell signaling, 12396S), FUNDC1 (From Quan Chen lab), FLAG (Sigma-Aldrich, F1804), HA (Sigma-Aldrich, H6533), ACTIN (Sigma-Aldrich, A2066), TOM70 (Proteintech, 14528-1-AP), TOM20 (ABclonal, A19403), SMAC (Cell signaling, 2954S), MITOFILIN (Proteintech, 10179-1-AP), HSP60 (Cell signaling, 4870S), TIMM23(Proteintech, 11123-1-AP), SKP1 (Cell signaling, 2156S), CUL1 (Proteintech, 12895-1-AP), Total OXPHOS Cocktail (Abcam, ab110411), Cytochrome C (BD Biosciences, 556433), mCherry (Easybio, BE2026), MTCH2 (Proteintech, 16888-1-AP), BECLIN1 (Cell signaling, 3495T), FIP200 (ABclonal, A14685), TFAM (Abcam, ab131607), LONP1 (Proteintech, 15440-1-AP), VDAC (Cell signaling, 4866S), Donkey polyclonal anti-Rabbit IgG (H+L),HRP-conjugated (Jackson ImmunoResearch, 711-035-152), Donkey polyclonal anti-Mouse IgG (H+L), HRP-conjugated (Jackson ImmunoResearch, 711-035-151), Goat polyclonal anti-Mouse IgG, Fcγ fragment specific, HRP-conjugated (Jackson ImmunoResearch, 115-035-008), Goat polyclonal anti-Mouse IgG, light chain specific, HRP-conjugated (Jackson ImmunoResearch, 115-035-174).

Techniques: Two Tailed Test, Western Blot