cleaved caspase 3  (Proteintech)

Journal: Materials Today Bio

Article Title: pH-responsive ZIF-8@quercetin nanoparticles induce pyroptosis for targeted gastric cancer therapy

doi: 10.1016/j.mtbio.2026.102806

Figure Lengend Snippet: Schematic illustration of the mechanism of ZIF-8@Que nanoparticles in GC treatment. ZIF-8@Que nanoparticles were prepared via a one-pot co-precipitation process, encapsulating quercetin within the ZIF-8 framework. Upon systemic administration, ZIF-8@Que preferentially accumulates in gastric tumors and undergoes acidic pH-triggered drug release. The nanoparticles exhibit intrinsic POD-like activity, catalyzing H 2 O 2 to generate ROS, which disrupt mitochondrial function by collapsing Δψm, depleting ATP, and enhancing oxidative stress. ROS-driven mitochondrial injury subsequently activates inflammasome-dependent caspase-1, leading to GSDMD cleavage, pore formation, release of IL-1β and IL-18, and subsequent pyroptotic cell death. This cascade culminates in robust eradication of GC cells while sparing normal tissues, highlighting the therapeutic promise of ZIF-8@Que as pyroptosis-inducing nanoparticles.

Article Snippet: Endogenous peroxidase was quenched with 3 % H 2 O 2 for 25 min. After blocking with 3 % BSA for 30 min, sections were incubated overnight at 4 °C with primary antibodies: Ki67 (1:500, CST), Cleaved caspase 3 (1:200, Abcam), Cleaved Caspase 1 (1:100, MCE), and Cleaved GSDMD (1:500, MCE).

Techniques: Activity Assay

Journal: Materials Today Bio

Article Title: pH-responsive ZIF-8@quercetin nanoparticles induce pyroptosis for targeted gastric cancer therapy

doi: 10.1016/j.mtbio.2026.102806

Figure Lengend Snippet: In vivo therapeutic efficacy and safety profile of ZIF-8@Que. (A) Hemolysis assay of mouse RBCs incubated with ZIF-8@Que at various concentrations. (B, C) Assessment of systemic toxicity in healthy mice: body weight (B) and serum biochemical (C) of C57BL/6 mice receiving repeated intravenous doses of ZIF-8@Que (50 mg/kg, once weekly for 4 weeks). (D) H&E staining of major organs confirming normal histology. Scale bar = 10 μm. (E) Experimental timeline for tumor establishment and treatment in nude mice. (F) Body weight of tumor-bearing mice during therapy. (G, H) Tumor growth curves (G) and tumor weight curves (H) for each group at different time points. (I) Kaplan-Meier survival curves for each group. (J) Representative tumor images at study endpoint. (K) H&E staining showing extensive necrosis in ZIF-8@Que-treated tumors. (L) Representative images of Ki67, cleaved caspase-1, cleaved caspase-3, cleaved GSDMD and TUNEL in tumor tissues, highlighting pyroptosis-mediated antitumor effects. For all studies, n ≥ 3. Data are shown as the mean ± SD. Comparisons were performed using the student's t-test or ANOVA. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, or ns = not significant.

Article Snippet: Endogenous peroxidase was quenched with 3 % H 2 O 2 for 25 min. After blocking with 3 % BSA for 30 min, sections were incubated overnight at 4 °C with primary antibodies: Ki67 (1:500, CST), Cleaved caspase 3 (1:200, Abcam), Cleaved Caspase 1 (1:100, MCE), and Cleaved GSDMD (1:500, MCE).

Techniques: In Vivo, Drug discovery, Hemolysis Assay, Incubation, Staining, TUNEL Assay

Journal: The FASEB Journal

Article Title: Timing of Glucocorticoid Treatment Dictates Glucocorticoid Receptor Actions Modulating the NLRP3‐Inflammasome Activation in Macrophages

doi: 10.1096/fj.202504083R

Figure Lengend Snippet: Glucocorticoids modulate inflammation and pyroptosis by regulating late NLRP3‐inflammasome activation through ACOD1 and iNOS. (A) Schematic representation of the experimental setup for conventional NLRP3‐inflammasome activation (upper), and late NLRP3‐inflammasome activation regulated by Dex and MCC950 (lower). (B) and (C) Quantification of IL‐1β concentration determined by ELISA in the supernatant of WT and GRKO BDMDs stimulated with LPS for 3 h (conventional) and 24 h (late), with Dex and vehicle added during the final 2 and 8 h of treatment, followed by ATP (5 mM) or Nigericin (10 μM) ( n = 3). (D) Immunoblot analysis depicting Dex effects on the protein levels of ACOD1, iNOS, NLRP3, pro‐IL‐1β, Caspase‐1 and GSDM‐D during late LPS‐induced NLRP3‐inflammasome activation and using ATP as signal 2 in WT and GRKO BMDMs lysates, detected with specific antibodies using western blotting. (E) Densitometry values of the immunoreactive bands quantified for protein content, normalized to β‐actin as depicted in (D) ( n = 4). (F) Nitrite levels in the supernatants of WT and GRKO BMDMs as depicted in (D) determined by colorimetric assay ( n = 4). (G) Assessment of lactate dehydrogenase (LDH) activity in the supernatants of WT and GRKO BMDMs as depicted in (D) ( n = 4). (H) Immunoblot analysis depicting the protein levels of NRF2 as depicted in (D) in presence of the proteasome inhibitor MG‐132 (2 μM) 6 h before harvesting cells ( n = 3). (I) Immunoblot analysis depicting the protein levels of HIF‐1α as depicted in (D) in presence of the prolyl‐hydroxylase inhibitor Roxadustat (RXD; 10 μM) 2 h before adding Dex ( n = 3). (J) Quantification of IL‐1β concentration determined by ELISA in the supernatant of WT BDMDs as depicted in (H) and (I) ( n = 3). (K) Assessment of lactate dehydrogenase (LDH) activity in supernatants of WT BMDMs subjected to the conditions described in (H) and (I) ( n = 3). Data represented as mean ± SEM. Blots shown are representative of a minimum of 3 independent experiments. Statistical analysis was performed using 2‐way ANOVA with Sidak's multiple‐comparison test (B–I); one‐way ANOVA with Tukey's multiple‐comparison test (J) and (K). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Lipopolysaccharide from E. coli 0111: B4 strain (LPS‐EB), NLRP3 inhibitor MCC950, Caspase‐1 inhibitor Ac‐Y‐VAD‐cmk, 4‐octyl‐itaconate (4‐OI), adenosine 5′‐triphosphate disodium salt (ATP), nigericin, and monosodium urate (MSU) crystals were purchased from InvivoGen (San Diego, CA, USA).

Techniques: Activation Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Colorimetric Assay, Activity Assay, Comparison

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