Journal: Redox Biology

Article Title: Redox-modulating macrophage biohybrid nanoplatform for targeted RIPK1-PANoptosome suppression in ischemic stroke

doi: 10.1016/j.redox.2025.103997

Figure Lengend Snippet: In vitro evaluation of microglia regulation of MA@ULips. (a) Scheme of microglia phenotype transition induced by MA@ULips. (b) Flow cytometry analysis of CD86 and CD206 in BV2 cells after diverse treatments. Representative immunofluorescence staining images of (c) CD86 and (d) CD206 in BV2 cells after various treatments. The level of (e) Arg-1, (f) IL-10, (g) TNF-α, and (h) IL-6 in the BV2 cells after different treatments (n = 3). Data are presented as mean ± SD. Statistical significance was calculated by one-way ANOVA with Tukey's multiple comparisons test. ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Anti-caspase 1 (31020-1-AP), anti-RIPK1 (17519-1-AP) and PE- anti -mouse CD86 + (PE-65068) were obtained from Proteintech Group.

Techniques: In Vitro, Flow Cytometry, Immunofluorescence, Staining

Journal: Redox Biology

Article Title: Redox-modulating macrophage biohybrid nanoplatform for targeted RIPK1-PANoptosome suppression in ischemic stroke

doi: 10.1016/j.redox.2025.103997

Figure Lengend Snippet: MA@ULips enhancing neurological functional recovery and modulating ischemic inflammatory microenvironment. (a) Experimental design involved multiple functional tests to evaluate the neurological outcomes of MA@ULips. (b) Forelimb asymmetry rate assessed through the cylinder test (n = 8). (c) Motion paths in the open field test. (d) Quantitative analysis of movement distance (n = 7). (e) Representative images of sham-operated mice and MCAO/R mice after different treatments. (f) Neurological deficit evaluation by the mNSS (n = 6). (g) Immunohistochemical staining for GFAP on brain tissue from the ischemic cortex area after different treatments. (h) Immunofluorescent staining of brain tissue for Iba-1 and CD86 after different treatments. (i–j) The levels of (i) IL-10 and (j) IL-6 in serum of ischemic mice after different treatments (n = 3). (k–l) The levels of (k) IL-6 and (l) IL-10 in brain tissues of ischemic mice after different treatments (n = 3). Data are presented as mean ± SD. Statistical significance was calculated by one-way ANOVA with Tukey's multiple comparisons test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Anti-caspase 1 (31020-1-AP), anti-RIPK1 (17519-1-AP) and PE- anti -mouse CD86 + (PE-65068) were obtained from Proteintech Group.

Techniques: Functional Assay, Immunohistochemical staining, Staining

Journal: iScience

Article Title: Size-dependent pyroptosis induction by calcium oxalate monohydrate crystals in HK-2 cells

doi: 10.1016/j.isci.2025.114459

Figure Lengend Snippet: Differences in pyroptosis induced by COM crystals of different sizes (A) Caspase-1/PI double staining flow cytometry quantitative analysis. (B) Caspase-1/PI/Hoechst 33342 triple staining confocal observation. Scale bars, 50 μm. (C) NLRP3 immunofluorescence map. Scale bars, 100 μm. (D) Quantitative histogram of pyroptosis; n = 3; mean ± SEM. (E) Quantitative bar graph of NLRP3; n = 3; mean ± SEM. NC: normal control. COM crystals were incubated with HK-2 cells for 48 h. Compared with NC group, ∗ p < 0.05; ∗∗ p < 0.01. (A) flow graph is divided into four regions (Q1, Q2, Q3, and Q4), of which Q1 is PI high signal area and Caspase-1 low signal area, representing apoptotic necrotic cells. Q2 is the PI high signal region and Caspase-1 high signal region, which represents the late pyroptosis cells. Q3 is the PI low signal area and Caspase-1 high signal area, which represents the early pyroptosis cells (Caspase-1 is actively expressed). Q4 is the PI hypointense region and Caspase-1 hypointense region, representing normal cells. Q1+ Q2 refers to dead cells, and Q2+ Q3 refers to pyroptosis cells. (B) shows the presence of four types of cells: the first type is the normal cell (indicated by the orange arrow), which corresponds to the Q4 region cells in (A); the second type was apoptotic or necrotic cells (indicated by white arrow), namely Q1 region cells. The third type was the late pyroptosis cells (yellow arrow), which were Q2 cells. The fourth type is the cells in the early stage of pyroptosis (purple arrow), which is the Q3 region cells. Orange arrows refer to normal cells, white arrows to apoptotic necrotic cells, yellow arrows to late pyroptosis cells, and purple arrows to early pyroptosis cells.

Article Snippet: Cleaved caspase-1 p20 , Proteintech , Cat No. 22915-1-AP; RRID: AB_2876874.

Techniques: Double Staining, Flow Cytometry, Staining, Immunofluorescence, Control, Incubation

Journal: iScience

Article Title: Size-dependent pyroptosis induction by calcium oxalate monohydrate crystals in HK-2 cells

doi: 10.1016/j.isci.2025.114459

Figure Lengend Snippet: Level of proteins in pyroptosis signaling pathway induced by COM crystals of different sizes (A) protein bands of NLRP3, cleaved caspase-1 p20, IL-18, and IL-1β. (B–E) Quantitative analysis of NLRP3, cleaved caspase-1 p20, IL-18, and IL-1β, respectively. n = 3; mean ± SEM. NC: normal control. COM crystals were incubated with HK-2 cells for 48 h.∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Cleaved caspase-1 p20 , Proteintech , Cat No. 22915-1-AP; RRID: AB_2876874.

Techniques: Control, Incubation

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 significantly inhibited OGD/R-induced pyroptosis in BV2 cells. A After transiently transfecting plasmids EGFP-N1-NLRP3, mCherry-C1-ASC, EGFP-N1-caspase-1, the cells underwent OGD/R with or without TA treatment, followed by Hoechst staining and imaging. Magnification: × 10; scale bar: 200 μm. B – D The bar chart presents the ratios of EGFP-NLRP3/Hoechst, mCherry-ASC/Hoechst, and EGFP-Caspase-1/Hoechst in different cell groups. ** p < 0.01 and *** p < 0.001 versus OGD/R alone group, n = 3. E Representative images of fluorescence expression following transiently transfecting with EGFP-GSDMD plasmid and counterstaining with DAPI. Magnification: 64 × , scale bar: 5 μm. F – I The Western blotting detection of the protein expressions of NLRP3, ASC, GSDMD, and IL-1β in BV2 cells subjected to OGD/R, with or without TA treatment. J – M The bar chart presents the ratios of NLRP3/GAPDH, ASC/GAPDH, N-GSDMD/GAPDH and Cleaved-IL-1β/GAPDH. * p < 0.05, ** p < 0.01 and *** p < 0.001versus OGD/R group, n = 3. N Representative images of Eth-D2/YO-PRO-1 staining in OGD/R-induced and TA or Triton-treated BV2 cells. Magnification: 10 × , scale bar: 200 μm. O The bar chart indicates the rate of YO-PRO-1 positivity cells. *** p < 0.001 versus OGD/R alone 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: Staining, Imaging, Fluorescence, Expressing, Plasmid Preparation, Western Blot

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