Journal: Frontiers in Endocrinology

Article Title: Spatio-temporal dynamics of autophagy-associated genes in macrophage-driven atherosclerosis: an integrated omics and experimental study

doi: 10.3389/fendo.2026.1764263

Figure Lengend Snippet: Expression detection of key genes in AS macrophages. (A) statistical graph of SNX5, SMG1, GSK3A mRNA expression level. (B) The SNX5, SMG1, GSK3A protein expression levels: Left, typical western blots, Right, statistical graph. (C) Protein expression of autophagy markers LC3 and p62. * P < 0.05, ** P < 0.01, *** P < 0.001 vs. Raw264.7.

Article Snippet: In addition, the protein expression levels of LC3 (Affinity, AF7001) and P62 (BIOSS, bs-8878R) were assessed to evaluate autophagic flux, with β-actin (Proteintech, 66009-1-Ig) serving as the loading control for normalization.

Techniques: Expressing, Western Blot

Journal: Poultry Science

Article Title: Ginsenoside 20(S)-Rg3 alleviates cage stress induced liver injury in laying ducks

doi: 10.1016/j.psj.2025.106254

Figure Lengend Snippet: Rg3 alleviated cage stress induced liver injury by suppressing inflammation and promoting autophagy. (A) Volcano plot (Rg3-H vs. ST); expression of inflammation-related genes in the liver of laying duck; IF of F4/80 protein in the liver of laying ducks. (B-C) Volcano plot (Rg3-H vs. ST); expression of antioxidant-related genes in the liver of laying duck; IF of GPX4 and ACSL4 protein in the liver of laying duck. (D) Volcano plot (Rg3-H vs. ST); expression of autophagy-related genes in the liver of laying duck; IF of LC3B protein in the liver of laying duck. Data are presented as the mean ± SEM; Intergroup differences were marked with lowercase letters. Different letters indicate statistically significant differences between groups (P < 0.05).

Article Snippet: Subsequently, 5 % bovine serum albumin (BSA) was added for blocking at room temperature for 1 h. Primary antibodies against F4/80 (28463-1-AP, Proteintech), GPX4 (30388-1-AP, Proteintech), ACSL4 (sc-365230, Santa Cruz), and LC3B (14600-1-AP, Proteintech) were added, and the sections were incubated overnight at 4°C.

Techniques: Expressing

Journal: Chinese Medicine

Article Title: Thonningianin A derived from Penthorum chinense Pursh alleviates cerebral ischemia/reperfusion-mediated apoptosis and pyroptosis through the activation of PINK1/Parkin-dependent mitophagy

doi: 10.1186/s13020-025-01247-2

Figure Lengend Snippet: TA activated mitophagy by PINK1/Parkin signaling pathway. A The Western blotting detection of the protein expressions of LC3I and LC3II in HT22 cells treated with TA and Rap (20 μM). B The bar chart presents the rate of LC3II/LC3I, * p < 0.05, * p < 0.01 and *** p < 0.01 versus the control group, n = 3. C Representative images of stable GFP-LC3 U87 cells treated with or without TA, Magnification: 20 × , scale bar: 100 μm. D The bar chart presents the rate of cells with GFP-LC3 puncta formation in stable GFP-LC3 U87 cells, * p < 0.05 and *** p < 0.001 versus control group. E Representative images of stable stable RFP-GFP-LC3 U87 cells treated with or without TA, Baf (100 nM) or Rap (10 μM), Magnification: × 64, scale bar: 5 μm. F Representative images of the co-localization of GFP-LC3 and MitoTracker in stable GFP-LC3 U87 cells with or without TA, CCCP or TA+AC220. G The bar chart indicates the rate of GFP/RFP, ** p < 0.05 and *** p < 0.001. H – K Line scan analysis showing the co-localization intensity (a.u.) of GFP-LC3 and MitoTracker in stable GFP-LC3 U87 cells under different treatments. L Representative images of HT22 cells transiently transfecting Mito-QC plasmid and treated with or without TA, CCCP or TA+AC220. M Representative images of BV2 cells transiently transfecting Mito-QC and treated with or without TA, CCCP or TA+AC220. N The bar chart indicates the rate of RFP/GFP in HT22 ransiently transfecting Mito-QC cells. ** p < 0.01 and *** p < 0.001 versus the control group, n = 3. O The bar chart indicates the rate of RFP/GFP in BV2 ransiently transfecting Mito-QC cells. ** p <0.01 and *** p < 0.001 versus the control group, n = 3. P Western blot detection of Parkin, p-Parkin, and PINK1 protein expression in HT22 cells treated with or without TA. Q , R The bar chart presents the rate of p-Parkin/Parkin (n = 3), PINK1/GAPDH (n = 5). ** p < 0.01 and *** p < 0.001 versus the control group

Article Snippet: Antibodies against GSDMD (20770-1-Ap), Caspase-3 (25158-1-Ap), Caspase-9 (10380-1-Ap), NQO1 (11451-1-Ap), GCLC (12601-1-Ap), LC3 (14600-1-Ap), PINK1 (23274-1-Ap), Parkin (14060-1-Ap), Antibody Caspase-1 (22,915-1-ap), GAPDH (6004-1-Ig), β-actin (66009-1-Ig), CoraLite ® Plus 488 goat anti-rabbit IgG (RGAR002), CoraLite ® Plus 594-Goat Anti-Mouse IgG (RGAR002), were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Western Blot, Control, Plasmid Preparation, Expressing

Journal: Chinese Medicine

Article Title: Thonningianin A derived from Penthorum chinense Pursh alleviates cerebral ischemia/reperfusion-mediated apoptosis and pyroptosis through the activation of PINK1/Parkin-dependent mitophagy

doi: 10.1186/s13020-025-01247-2

Figure Lengend Snippet: TA inhibited OGD/R-induced pyroptosis by activating mitophagy in BV2 cells. A BV2 cells were subjected to OGD/R and treated with or wthout TA, then cell viability were detected by MTT.*** p < 0.001 versus OGD/R alone group, n = 3. B BV2 cells were subjected to OGD/R and treated either TA or TA combined with mitophagy inhibitor AC220, then cell viability was detected by MTT. ** p < 0.01 and *** p < 0.001 versus the corresponding group, n = 3. C Representative images of Hoechst and PI staining in BV2 cells subjected to OGD/R and treated with either TA or TA combined with AC220. Magnification: × 10; scale bar: 100 μm. D The Western blotting detection of the protein expressions of NLRP3 and GSDMD in BV2 cells subjected to OGD/R and treated with either TA or TA combined with atophagy inhibitor 3MA. E The bar chart indicates the rate of PI/Hoechst in BV2 cells. *** p < 0.001, n = 3. F , G The bar chart presents the ratios of NLRP3/β-actin, N-GSDMD/β-actin. * p < 0.05 and *** p < 0.001versus the corresponding group, n = 3. H Representative images of BV2 cells transfected with EGFP-NLRP3, EGFP-Caspase-1 or mCherry-ASC plasmid and subjected to OGD/R and treated with either TA or TA combined with AC220. Magnification: × 10, scale bar: 100 μm. I BV2 cells were subjected to OGD/R and treated with or without TA, immunofluorescence staining was used to observe the co-localization of LC3 and NLRP3, and fluorescence images were collected. Magnification: × 64, scale bar: 5 μm. J – L The bar chart presents the ratios of EGFP-NLRP3/Hoechst, EGFP-Caspase-1/Hoechst or mCherry-ASC/Hoechst in BV2 cells of different groups. * p < 0.05, ** p < 0.01 and *** p < 0.001, n = 3

Article Snippet: Antibodies against GSDMD (20770-1-Ap), Caspase-3 (25158-1-Ap), Caspase-9 (10380-1-Ap), NQO1 (11451-1-Ap), GCLC (12601-1-Ap), LC3 (14600-1-Ap), PINK1 (23274-1-Ap), Parkin (14060-1-Ap), Antibody Caspase-1 (22,915-1-ap), GAPDH (6004-1-Ig), β-actin (66009-1-Ig), CoraLite ® Plus 488 goat anti-rabbit IgG (RGAR002), CoraLite ® Plus 594-Goat Anti-Mouse IgG (RGAR002), were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Staining, Western Blot, Transfection, Plasmid Preparation, Immunofluorescence, Fluorescence

Journal: Chinese Medicine

Article Title: Thonningianin A derived from Penthorum chinense Pursh alleviates cerebral ischemia/reperfusion-mediated apoptosis and pyroptosis through the activation of PINK1/Parkin-dependent mitophagy

doi: 10.1186/s13020-025-01247-2

Figure Lengend Snippet: TA ameliorated neurological injury in MCAO/R Rats. A Representative immunofluorescence staining images of LC3 and NLRP3 in hippocampus of the brain in MCAO/R rats, × 10, scale bar: 100 μm. B The bar chart presents the counts of LC3 + cells, NLRP3 + /cells, and LC3 + /NLRP3 + co-localized cells. C – F The Western blotting detection of the protein expressions of LC3, PINK1, p-Parkin and NLRP3 in rat brain tissues across experimental groups. G – J The bar chart presents the ratios of LC3-II/I, PINK1/β-actin, p-Parkin/ Parkin, NLRP3/β-actin. * p < 0.05, ** p < 0.01 and *** p < 0.001versus the alone MCAO/R group, n = 3. K – N The Western blotting detection of the protein expressions of ASC, Caspase-1, IL-18 in rat brain tissues across experimental groups. O – R The bar chart presents the ratios of ASC/β-actin, Cleaved-Caspase-1/Pro- Caspase-1, Cleaved-IL-18/β-actin. * p < 0.05, ** p < 0.01 and *** p < 0.001versus the alone MCAO/R group, n = 3. S The Western blotting detection of the protein expression of IL-1β. T The bar chart presents the rate of Cleaved-IL-18/GAPDH. * p < 0.05 and ** p < 0.01 versus the alone MCAO/R group, n = 3. S The Western blotting detection of the protein expression of IL-1β. T The bar chart presents the rate of Cleaved-IL-1β/GAPDH. * p < 0.05 and ** p < 0.01 versus the alone MCAO/R group, n = 3. U The Western blotting detection of the protein expression of BAX and Bcl2. V The bar chart presents the rate of BAX/Bcl2. *** p < 0.001 versus the alone MCAO/R group, n = 3

Article Snippet: Antibodies against GSDMD (20770-1-Ap), Caspase-3 (25158-1-Ap), Caspase-9 (10380-1-Ap), NQO1 (11451-1-Ap), GCLC (12601-1-Ap), LC3 (14600-1-Ap), PINK1 (23274-1-Ap), Parkin (14060-1-Ap), Antibody Caspase-1 (22,915-1-ap), GAPDH (6004-1-Ig), β-actin (66009-1-Ig), CoraLite ® Plus 488 goat anti-rabbit IgG (RGAR002), CoraLite ® Plus 594-Goat Anti-Mouse IgG (RGAR002), were purchased from Proteintech Group, Inc (Wuhan, China).

Techniques: Immunofluorescence, Staining, Western Blot, Expressing