nlrp3 ko mice  (Jackson Laboratory)

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Propranolol suppressed NLRP3 inflammasome activation in tMCAO mice (A) Western blot analysis of IBA-1, NLRP3, and IL-1β in the ischemic penumbra of different groups. (B–D) Quantitative analysis of IBA-1, NLRP3, and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the Sham-operated group; # p < 0.05 versus the tMCAO group. (E) Immunofluorescence staining for NLRP3 of brain sections in the indicated groups. Scale bars, 100 μm (main image) and 30 μm (magnified view). (F) Quantitative analysis of NLRP3 positive cells ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the Sham-operated group; # p < 0.05 versus the tMCAO group. (G) Immunofluorescence staining for IBA-1 of brain sections in the indicated groups. Scale bars, 50 μm (main image) and 10 μm (magnified view). (H) Representative images of microglia and heat maps of Sholl analysis were shown. Scale bar, 10 μm. (I) Sholl profile shows the number of intersections at different distances from the microglial soma in the indicated groups ( n = 4 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using two-way ANOVA followed by Tukey’s post hoc test. ∗∗∗∗ p < 0.0001 versus the Sham-operated group; #### p < 0.0001 versus the tMCAO group. (J and K) Quantitative analysis of process length (J) and endpoints (K) per cell ( n = 4 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the Sham-operated group; # p < 0.05and ## p < 0.01 versus the tMCAO group.

Article Snippet: NLRP3 KO mice (male, 8-12 weeks old) , Jackson Laboratory , Strain #021302.

Techniques: Activation Assay, Western Blot, Expressing, Immunofluorescence, Staining

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: NLRP3 -/- mice exhibited reduced cerebral infarction volume and attenuated BBB damage (A) TTC staining was performed to measure the cerebral infarct volume in mice. Scale bar, 5 mm. (B) Statistical analysis of infarction volume in different groups ( n = 6 mice/group). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ## p < 0.01 versus the WT tMCAO group; ns, not significant. (C) Neurological function evaluation was conducted using the mNSS score ( n = 8 mice/group). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. # p < 0.05 versus the WT tMCAO group; ns, not significant. (D) Western blot analysis of NLRP3, IL-1β, Occludin, and ZO-1 in the ischemic penumbra of different groups. (E) Quantitative analysis of NLRP3, IL-1β, Occludin, and ZO-1 expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 versus the WT Sham-operated group; # p < 0.05, ## p < 0.01, and #### p < 0.0001 versus the WT tMCAO group; ns, not significant. (F) Representative images of brain slices show the extravasation of EB. Scale bar, 5 mm. (G) Quantitative analysis of EB leakage in mice from different groups ( n = 4 mice/group). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. # p < 0.05 versus the WT tMCAO group; ns, not significant.

Article Snippet: NLRP3 KO mice (male, 8-12 weeks old) , Jackson Laboratory , Strain #021302.

Techniques: Staining, Western Blot, Expressing

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Knockout of NLRP3 alleviated BBB damage and the activation of microglia caused by tMCAO (A) Immunofluorescence staining for IBA-1 of brain sections in the indicated groups. Scale bars, 50 μm (main image) and 10 μm (magnified view). (B) Representative images of microglia and heat maps of Sholl analysis were shown. Scale bar, 10 μm. (C) Sholl profile shows the number of intersections at different distances from the microglial soma in the indicated groups ( n = 4 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using two-way ANOVA followed by Tukey’s post hoc test. ∗∗∗∗ p < 0.0001 versus the WT Sham-operated group; #### p < 0.0001 versus the WT tMCAO group. (D and E) Quantitative analysis of process length (D) and endpoints (E) per cell ( n = 4 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the WT Sham-operated group; # p < 0.05 versus the WT tMCAO group; ns, not significant. (F) Immunofluorescence staining for CD-31 of brain sections of the indicated groups. Scale bars, 100 μm (main image) and 30 μm (magnified view). (G) Statistical analysis of CD-31 positive cells ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the WT Sham-operated group; # p < 0.05 versus the WT tMCAO group; ns, not significant.

Article Snippet: NLRP3 KO mice (male, 8-12 weeks old) , Jackson Laboratory , Strain #021302.

Techniques: Knock-Out, Activation Assay, Immunofluorescence, Staining

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Propranolol reduced the expression of NLRP3 and IL-1β in BV2 cells subjected to OGD/R (A) Schematic of cell experiment steps. (B) Western blot analysis of NLRP3 and IL-1β in BV2 cells at 4, 6, 8, 12, and 24 h after OGD/R. (C and D) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 versus the control group. (E) Western blot analysis of NLRP3 and IL-1β in BV2 cells of different treatment groups. (F and G) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 and ## p < 0.01 versus the OGD/R group.

Article Snippet: NLRP3 KO mice (male, 8-12 weeks old) , Jackson Laboratory , Strain #021302.

Techniques: Expressing, Western Blot, Control

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Blockade of β2-AR reduced the expression of NLRP3 in BV2 cells subjected to OGD/R (A) Western blot analysis of the effects of dobutamine and betaxolol on NLRP3 and IL-1β expression in BV2 cells. (B and C) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the control group. (D) Western blot analysis of salmeterol and ICI118,551 on NLRP3 and IL-1β expression in BV2 cells. (E and F) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 and ## p < 0.01 versus the OGD/R group.

Article Snippet: NLRP3 KO mice (male, 8-12 weeks old) , Jackson Laboratory , Strain #021302.

Techniques: Expressing, Western Blot, Control

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Activation of β2-AR induced the expression of NLRP3 and IL-1β via the ERK1/2 MAPK signaling pathway (A) Western blot analysis of the effects of H-89 and U0126 on NLRP3 and IL-1β expression in BV2 cells. (B and C) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 versus the OGD/R group. (D) Western blot analysis of p -ERK1/2 expression in BV2 cells. (E) Quantitative analysis of p -ERK1/2 expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 versus the OGD/R group.

Article Snippet: NLRP3 KO mice (male, 8-12 weeks old) , Jackson Laboratory , Strain #021302.

Techniques: Activation Assay, Expressing, Western Blot, Control

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Propranolol suppressed NLRP3 inflammasome activation in tMCAO mice (A) Western blot analysis of IBA-1, NLRP3, and IL-1β in the ischemic penumbra of different groups. (B–D) Quantitative analysis of IBA-1, NLRP3, and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the Sham-operated group; # p < 0.05 versus the tMCAO group. (E) Immunofluorescence staining for NLRP3 of brain sections in the indicated groups. Scale bars, 100 μm (main image) and 30 μm (magnified view). (F) Quantitative analysis of NLRP3 positive cells ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the Sham-operated group; # p < 0.05 versus the tMCAO group. (G) Immunofluorescence staining for IBA-1 of brain sections in the indicated groups. Scale bars, 50 μm (main image) and 10 μm (magnified view). (H) Representative images of microglia and heat maps of Sholl analysis were shown. Scale bar, 10 μm. (I) Sholl profile shows the number of intersections at different distances from the microglial soma in the indicated groups ( n = 4 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using two-way ANOVA followed by Tukey’s post hoc test. ∗∗∗∗ p < 0.0001 versus the Sham-operated group; #### p < 0.0001 versus the tMCAO group. (J and K) Quantitative analysis of process length (J) and endpoints (K) per cell ( n = 4 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the Sham-operated group; # p < 0.05and ## p < 0.01 versus the tMCAO group.

Article Snippet: Male C57BL/6J mice aged 8-12 weeks were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China), while NLRP3 KO (NLRP3 -/- ) mice with C57BL/6 background were purchased from Jackson Laboratory.

Techniques: Activation Assay, Western Blot, Expressing, Immunofluorescence, Staining

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: NLRP3 -/- mice exhibited reduced cerebral infarction volume and attenuated BBB damage (A) TTC staining was performed to measure the cerebral infarct volume in mice. Scale bar, 5 mm. (B) Statistical analysis of infarction volume in different groups ( n = 6 mice/group). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ## p < 0.01 versus the WT tMCAO group; ns, not significant. (C) Neurological function evaluation was conducted using the mNSS score ( n = 8 mice/group). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. # p < 0.05 versus the WT tMCAO group; ns, not significant. (D) Western blot analysis of NLRP3, IL-1β, Occludin, and ZO-1 in the ischemic penumbra of different groups. (E) Quantitative analysis of NLRP3, IL-1β, Occludin, and ZO-1 expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 versus the WT Sham-operated group; # p < 0.05, ## p < 0.01, and #### p < 0.0001 versus the WT tMCAO group; ns, not significant. (F) Representative images of brain slices show the extravasation of EB. Scale bar, 5 mm. (G) Quantitative analysis of EB leakage in mice from different groups ( n = 4 mice/group). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. # p < 0.05 versus the WT tMCAO group; ns, not significant.

Article Snippet: Male C57BL/6J mice aged 8-12 weeks were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China), while NLRP3 KO (NLRP3 -/- ) mice with C57BL/6 background were purchased from Jackson Laboratory.

Techniques: Staining, Western Blot, Expressing

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Knockout of NLRP3 alleviated BBB damage and the activation of microglia caused by tMCAO (A) Immunofluorescence staining for IBA-1 of brain sections in the indicated groups. Scale bars, 50 μm (main image) and 10 μm (magnified view). (B) Representative images of microglia and heat maps of Sholl analysis were shown. Scale bar, 10 μm. (C) Sholl profile shows the number of intersections at different distances from the microglial soma in the indicated groups ( n = 4 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using two-way ANOVA followed by Tukey’s post hoc test. ∗∗∗∗ p < 0.0001 versus the WT Sham-operated group; #### p < 0.0001 versus the WT tMCAO group. (D and E) Quantitative analysis of process length (D) and endpoints (E) per cell ( n = 4 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the WT Sham-operated group; # p < 0.05 versus the WT tMCAO group; ns, not significant. (F) Immunofluorescence staining for CD-31 of brain sections of the indicated groups. Scale bars, 100 μm (main image) and 30 μm (magnified view). (G) Statistical analysis of CD-31 positive cells ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the WT Sham-operated group; # p < 0.05 versus the WT tMCAO group; ns, not significant.

Article Snippet: Male C57BL/6J mice aged 8-12 weeks were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China), while NLRP3 KO (NLRP3 -/- ) mice with C57BL/6 background were purchased from Jackson Laboratory.

Techniques: Knock-Out, Activation Assay, Immunofluorescence, Staining

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Propranolol reduced the expression of NLRP3 and IL-1β in BV2 cells subjected to OGD/R (A) Schematic of cell experiment steps. (B) Western blot analysis of NLRP3 and IL-1β in BV2 cells at 4, 6, 8, 12, and 24 h after OGD/R. (C and D) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001 versus the control group. (E) Western blot analysis of NLRP3 and IL-1β in BV2 cells of different treatment groups. (F and G) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 and ## p < 0.01 versus the OGD/R group.

Article Snippet: Male C57BL/6J mice aged 8-12 weeks were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China), while NLRP3 KO (NLRP3 -/- ) mice with C57BL/6 background were purchased from Jackson Laboratory.

Techniques: Expressing, Western Blot, Control

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Blockade of β2-AR reduced the expression of NLRP3 in BV2 cells subjected to OGD/R (A) Western blot analysis of the effects of dobutamine and betaxolol on NLRP3 and IL-1β expression in BV2 cells. (B and C) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < 0.001 versus the control group. (D) Western blot analysis of salmeterol and ICI118,551 on NLRP3 and IL-1β expression in BV2 cells. (E and F) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 and ## p < 0.01 versus the OGD/R group.

Article Snippet: Male C57BL/6J mice aged 8-12 weeks were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China), while NLRP3 KO (NLRP3 -/- ) mice with C57BL/6 background were purchased from Jackson Laboratory.

Techniques: Expressing, Western Blot, Control

Journal: iScience

Article Title: Propranolol alleviates cerebral infarction through the β2-AR-mediated ERK/NLRP3 pathway

doi: 10.1016/j.isci.2026.115779

Figure Lengend Snippet: Activation of β2-AR induced the expression of NLRP3 and IL-1β via the ERK1/2 MAPK signaling pathway (A) Western blot analysis of the effects of H-89 and U0126 on NLRP3 and IL-1β expression in BV2 cells. (B and C) Quantitative analysis of NLRP3 and IL-1β expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 versus the OGD/R group. (D) Western blot analysis of p -ERK1/2 expression in BV2 cells. (E) Quantitative analysis of p -ERK1/2 expression ( n = 3 biological replicates per group, each with 3 technical replicates). Data were expressed as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus the control group; # p < 0.05 versus the OGD/R group.

Article Snippet: Male C57BL/6J mice aged 8-12 weeks were purchased from the Model Animal Research Center of Nanjing University (Nanjing, China), while NLRP3 KO (NLRP3 -/- ) mice with C57BL/6 background were purchased from Jackson Laboratory.

Techniques: Activation Assay, Expressing, Western Blot, Control

Journal: Science advances

Article Title: The P2X7R/NLRP3 inflammasome axis suppresses enthesis regeneration through inflammatory and metabolic macrophage-stem cell cross-talk.

doi: 10.1126/sciadv.adr4894

Figure Lengend Snippet: Fig. 1. NLRP3 inflammasomes are activated in macrophages at the injured enthesis. (A) Schematic diagram of RNA-seq and validation of DEGs. (B) PCA of the sham operation and RCTR groups. (C) Heat map of DEGs between the sham operation and RCTR groups. (D) Relative mRNA expression levels of Il1b, Caspase-1, Nlrp3, and P2rx7 in the enthesis of RCTR groups in comparison to sham operation groups. (E) Immunofluorescence (IF) staining of CD68 (red), NLRP3 (green), and caspase-1 (magenta) in the injured enthesis of wild-type and Nlrp3−/− mice at 3 dpi. Orange and green dashed squares represent enlarged images of the enthesis. Arrows indicate specks of NLRP3 and caspase-1. (F) Quantification of specks per macrophage in the injured enthesis of wild-type and Nlrp3−/− mice at 3 dpi. (G and H) The relative caspase-1 activity and IL-1β concentration of the enthesis in wild-type and Nlrp3−/− mice at 3, 7, 14, and 28 dpi. T, tendon; I, tendon-to-bone interface; B, bone. Data are presented as means ± SD. Statistical significance was determined using one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test. d, days.

Article Snippet: Nlrp3 KO (Nlrp3−/−) mice (stock no. S- KO- 05210) and P2rx7 KO (P2rx7 KO) mice (stock no. S- KO- 03541) were obtained from Cyagen (Suzhou, China).

Techniques: RNA Sequencing, Biomarker Discovery, Expressing, Comparison, Immunofluorescence, Staining, Activity Assay, Concentration Assay

Journal: Science advances

Article Title: The P2X7R/NLRP3 inflammasome axis suppresses enthesis regeneration through inflammatory and metabolic macrophage-stem cell cross-talk.

doi: 10.1126/sciadv.adr4894

Figure Lengend Snippet: Fig. 2. The activation of NLRP3 inflammasomes inhibits enthesis regeneration. (A) Schematic of animal experiments, in which the enthesis injury and the repair model were established in wild-type and Nlrp3−/− mice, with analyses at 3, 7, 14, and 28 dpi. (B) H&E and toluidine blue staining of the enthesis in wild-type and Nlrp3−/− mice at 14 and 28 dpi. Black dashed squares represent enlarged images of the enthesis. (C and D) Metachromasia area size and histological scores of the enthesis in wild- type and Nlrp3−/− mice at 14 and 28 dpi. (E) Micro-CT coronal views of the humerus of wild-type and Nlrp3−/− mice at 14 and 28 dpi. Green dashed squares represent the area of the enthesis. (F to H) Quantitative analysis of BMD, BV/TV, and Tb.N of the enthesis. (I) Deformation and load curves of the enthesis in wild-type and Nlrp3−/− mice at 14 and 28 dpi. (J to L) Failure load, stiffness, and work of the enthesis in wild-type and Nlrp3−/− mice at 14 and 28 dpi. Data are presented as means ± SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test and Student’s t test.

Article Snippet: Nlrp3 KO (Nlrp3−/−) mice (stock no. S- KO- 05210) and P2rx7 KO (P2rx7 KO) mice (stock no. S- KO- 03541) were obtained from Cyagen (Suzhou, China).

Techniques: Activation Assay, Staining, Micro-CT

Journal: Science advances

Article Title: The P2X7R/NLRP3 inflammasome axis suppresses enthesis regeneration through inflammatory and metabolic macrophage-stem cell cross-talk.

doi: 10.1126/sciadv.adr4894

Figure Lengend Snippet: Fig. 3. scRNA-seq uncovers that NLRP3 inflammasomes deteriorate inflammation and IL-1β inflammatory cross-talk. (A) Schematic of scRNA-seq, in which the enthesis was harvested from wild-type controls and Nlrp3−/− mice at 7 dpi and processed for scRNA-seq. (B) UMAP plot of 56,217 cells from wild-type controls (n = 3) and Nlrp3−/− mice (n = 3). (C) Bar plot of the proportions of nine major cell clusters in the injured enthesis of wild-type controls and Nlrp3−/− mice at 7 dpi. (D) Violin plots of specific gene expressions in macrophages, mesenchymal cells, and neutrophils. (E) GO enrichment analysis of down-regulated genes in PIM and up-regulated genes in AIM in Nlrp3−/− mice. (F) Circle plot of the interactions of subsets of macrophages and mesenchymal cells. Edge line thickness suggests the interaction strength between different cell clusters. (G) NicheNet analysis of ligand-target regulatory potential between macrophages and mesenchymal stem cells. (H) Gene set cores of IL-1 signaling in different mesenchymal cell subsets in wild-type controls and Nlrp3−/− mice. Statistical significance was determined using Student’s t test. FACS, fluorescence-activated cell sorting; TGF-β, transforming growth factor–β.

Article Snippet: Nlrp3 KO (Nlrp3−/−) mice (stock no. S- KO- 05210) and P2rx7 KO (P2rx7 KO) mice (stock no. S- KO- 03541) were obtained from Cyagen (Suzhou, China).

Techniques: Fluorescence, FACS

Journal: Science advances

Article Title: The P2X7R/NLRP3 inflammasome axis suppresses enthesis regeneration through inflammatory and metabolic macrophage-stem cell cross-talk.

doi: 10.1126/sciadv.adr4894

Figure Lengend Snippet: Fig. 5. NLRP3 inflammasomes suppress the secretion of anti-inflammatory cytokines by macrophages to inhibit inflammation resolution. (A) IL-1β concentration in the supernatant of wild-type and Nlrp3−/− BMDMs. (B) Scan images of the supernatant of wild-type and Nlrp3−/− BMDMs in the mouse inflammation array Q1. (C) Heat- map of inflammation factors in the supernatant of wild-type and Nlrp3−/− BMDMs. (D) Concentrations of IL-1β, IFN-γ, IL-6, TNF-α, IL-1α, IL-10, IL-13, and IL-4 in the super- natant of wild-type and Nlrp3−/− BMDMs. (E to H) IHC staining and ELISA analysis of IL-10 and IL-13 in the enthesis of wild-type and Nlrp3−/− mice. Green dashed squares represent the enlarged images of the enthesis. (I and J) Immunofluorescence staining and quantification of CD68 (green)– and CD206 (red)–positive cells in the enthesis of wild-type controls and Nlrp3−/− mice. Red dashed squares represent the enlarged images of the enthesis. Arrows indicate CD68+ and CD206+ macrophages. Data are presented as means ± SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test and Student’s t test.

Article Snippet: Nlrp3 KO (Nlrp3−/−) mice (stock no. S- KO- 05210) and P2rx7 KO (P2rx7 KO) mice (stock no. S- KO- 03541) were obtained from Cyagen (Suzhou, China).

Techniques: Concentration Assay, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining

Journal: Science advances

Article Title: The P2X7R/NLRP3 inflammasome axis suppresses enthesis regeneration through inflammatory and metabolic macrophage-stem cell cross-talk.

doi: 10.1126/sciadv.adr4894

Figure Lengend Snippet: Fig. 6. NLRP3 inflammasomes inhibit the production of docosatrienoic acid. (A) Schematic of untargeted metabolomics of EIF. (B) PCA of untargeted metabolomics of EIF. (C) The number of differential metabolites in EIF. (D) Schematic of untargeted metabolomics and RNA sequencing of wild-type and Nlrp3−/− BMDMs. (E) PCA of untargeted metabolomics of supernatant from wild-type and Nlrp3−/− BMDMs. (F) The number of differential metabolites in the supernatant. (G) Venn diagram showing significantly enriched metabolites in different experimental setting. (H and I) Mass spectrometer quantification of DTA in EIF and supernatant. (J) PCA of RNA sequencing of wild-type and Nlrp3−/− BMDMs. (K) GO enrichment analysis of differentially expressed pathways of BMDMs. (L) Gene set enrichment analysis (GSEA) of biosynthesis of unsaturated fatty acids. (M) Gene set cores of unsaturated fatty acid biosynthetic process and regulation of unsaturated fatty acid biosynthetic process in PIM and AIM in wild-type controls and Nlrp3−/− mice. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test and Student’s t test.

Article Snippet: Nlrp3 KO (Nlrp3−/−) mice (stock no. S- KO- 05210) and P2rx7 KO (P2rx7 KO) mice (stock no. S- KO- 03541) were obtained from Cyagen (Suzhou, China).

Techniques: RNA Sequencing, Mass Spectrometry

Journal: Science advances

Article Title: The P2X7R/NLRP3 inflammasome axis suppresses enthesis regeneration through inflammatory and metabolic macrophage-stem cell cross-talk.

doi: 10.1126/sciadv.adr4894

Figure Lengend Snippet: Fig. 8. Conditional KO of P2rx7 in myeloid cells reduces NLRP3 inflammasome activation after enthesis injury and improves enthesis regeneration. (A) Immuno- fluorescence staining of CD68 (green) and P2X7R (yellow) in native and injured enthesis. Red dashed squares represent enlarged images of the enthesis. (B) Feature plots of single-cell gene expression of P2rx7 in macrophages in wild-type mice. (C and D) The relative caspase-1 activity and concentration of IL-1β in the enthesis of Lyz2-P2rx7f/f and Lyz2-P2rx7−/− mice at 3, 7, 14, and 28 dpi. (E) Schematic of animal experiments, in which the RCTR model was established in Lyz2-P2rx7f/f and Lyz2-P2rx7−/− mice, and analyzed at 3, 7, 14, and 28 dpi. (F) H&E and toluidine blue staining of the enthesis in Lyz2-P2rx7f/f and Lyz2-P2rx7−/− mice at 14 and 28 dpi. Black dashed squares represent enlarged images of the enthesis. (G and H) Histological scores and the metachromasia area size of the enthesis in Lyz2-P2rx7f/f and Lyz2-P2rx7−/− mice at 14 and 28 dpi. (I) Micro-CT coronal views of the humerus of Lyz2-P2rx7f/f and Lyz2-P2rx7−/− mice at 14 and 28 dpi. Green dashed squares represent the area of interest. (J and K) Quantitative analysis of the BMD and BV/TV of the enthesis. (L) Deformation and load curves of the enthesis in Lyz2-P2rx7 f/f and Lyz2-P2rx7−/− mice at 14 and 28 dpi. (M to O) Failure load, stiffness, and work of the enthesis in mice with IL-1β neutralizing antibodies or control IgG injection at 14 and 28 dpi. Data are presented as means ± SD. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test and Student’s t test.

Article Snippet: Nlrp3 KO (Nlrp3−/−) mice (stock no. S- KO- 05210) and P2rx7 KO (P2rx7 KO) mice (stock no. S- KO- 03541) were obtained from Cyagen (Suzhou, China).

Techniques: Activation Assay, Fluorescence, Staining, Gene Expression, Activity Assay, Concentration Assay, Micro-CT, Control, Injection

Journal: Science advances

Article Title: The P2X7R/NLRP3 inflammasome axis suppresses enthesis regeneration through inflammatory and metabolic macrophage-stem cell cross-talk.

doi: 10.1126/sciadv.adr4894

Figure Lengend Snippet: Fig. 9. The schematic of this study. After enthesis injury, NLRP3 inflammasomes are activated in infiltrated macrophages upon receiving activation signals mediated by P2X7R. The activation of the P2X7R/NLRP3 inflammasome axis not only exacerbates inflammation by prompting the release of IL-1β and suppressing the production of anti-inflammatory factors including IL-10 and IL-13 but also inhibits the production of proregenerative docosatrienoic acid. NLRP3 inflammasomes suppress enthesis re- generation via aggravating IL-1β inflammatory cross-talk and restraining docosatrienoic acid metabolic cross-talk between macrophages and stem cells. Blocking the P2X7R/NLRP3 inflammasome axis rewires the cross-talk between macrophages and stem cells and converts pathological inflammation to reparative inflammation. This study illustrates that the P2X7R/NLRP3 inflammasome axis is a promising regenerative therapeutic target for enthesis injury treatment.

Article Snippet: Nlrp3 KO (Nlrp3−/−) mice (stock no. S- KO- 05210) and P2rx7 KO (P2rx7 KO) mice (stock no. S- KO- 03541) were obtained from Cyagen (Suzhou, China).

Techniques: Activation Assay, Blocking Assay

Journal: Journal of Translational Medicine

Article Title: Targeting the NLRP3 in macrophages contributes to senescence cell clearance in radiation-induced skin injury

doi: 10.1186/s12967-025-06204-z

Figure Lengend Snippet: Macrophages with high senescence clearance capacity upregulate the expression of Nlrp3. A DEGs were analyzed using a log2-fold change expression and the percentage difference in cells expressing the gene, for macrophages with low and high senescence clearance capacity (Δ percentage difference). The log2-fold change of gene was > 1, and the adjusted P value was < 0.05 by the Wilcoxon rank sum test. B Enrichment of KEGG pathways for DEGs in macrophages with low and high senescence clearance capacity. C Protein interaction network and hub genes of DEGs expressed in macrophages with high senescence clearance capacity. D Violin plot showing the gene expression level of Nlrp3 in macrophages at different time points after radiation. E Western blot of NLRP3, CASPASE1, and IL-18 protein expression levels in skin tissue at different time points after radiation. F RT-qPCR analysis of Nlrp3 gene expression levels in skin tissue at different time points after radiation, n = 3, one-way ANOVA with Tukey HSD post hoc comparison. G Representative images of multiplex immunofluorescence for F4/80, NLRP3, P21 at different time points after radiation. Scale bar, 100, 20 μm. H Dimensionality reduction shows the definition and marker genes of the 5 macrophage subsets. I Characteristics of the distribution of macrophage clusters with low and high senescence cell clearance capabilities. DEGs, differentially expressed genes. KEGG, kyoto encyclopedia of genes and genomes. Data were represented as the mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The C57BL/6J mice and SD rats were obtained from the Animal Center of Army Medical University, NLRP3 flox/flox mice and Lyz2-Cre mice were constructed by Cyagen (GuangZhou, China) and used to generate macrophage-specific Nlrp3 conditional knockout mice (NLRP3 Lyz−/− ).

Techniques: Expressing, Gene Expression, Western Blot, Quantitative RT-PCR, Comparison, Multiplex Assay, Immunofluorescence, Marker

Journal: Journal of Translational Medicine

Article Title: Targeting the NLRP3 in macrophages contributes to senescence cell clearance in radiation-induced skin injury

doi: 10.1186/s12967-025-06204-z

Figure Lengend Snippet: NLRP3 mediates the clearance of senescent fibroblasts by macrophages. A Schematic diagram of treatment of in situ macrophages in 20-day post-radiation skin tissue with NLRP3 agonists and inhibitors. B Western blot of NLRP3, CASPASE1, and IL-18 protein levels in in situ macrophages treated with NLRP3agonist and inhibitor. C RT-qPCR analysis of Il18 , Il1b , Caspase1 , and Aim2 gene expression levels in in situ macrophages treated with NLRP3 agonist and inhibitor, n = 3, one-way ANOVA with Tukey HSD post hoc comparison. D Representative images and statistics of Transwell assays after NLRP3 agonist and inhibitor treatment of in situ macrophages, n = 3, one-way ANOVA with Tukey HSD post hoc comparison,Scale bar 50 μm. E Representative images and statistics of the engulfment capacity after treatment of in situ macrophages with NLRP3 agonists and inhibitors, n = 3, one-way ANOVA,Scale bar 50 μm. F Schematic diagram of the co-culture of in situ macrophages and senescent fibroblasts after treatment with NLRP3 agonists or inhibitors. G Representative images of senescence cells co-cultured with differently treated macrophages were imaged within 24 h, showing the same field of view over time. Scale bar, 50 μm. H Quantification of senescence cell green area signal at different time points after co-culture, n = 3, two-way ANOVA . I Western blot of NLRP3 protein expression levels in in situ macrophages from WT mouse and Nlrp3 Lyz−/− mouse. J Evaluation of the phagocytic capacity in in situ macrophages from WT mouse and Nlrp3 Lyz−/− mouse, n = 3, t test. K , L Flow cytometry assessment of senescence cell viability after co-culture of in situ macrophages from WT mouse and Nlrp3 Lyz−/− mouse with senescent fibroblasts, n = 3, one-way ANOVA with Tukey HSD post hoc comparison. M Representative images of senescence cells co-cultured with in situ macrophages from WT mouse or Nlrp3 Lyz−/− mouse for 24 h, showing the same field over time. Scale bar, 50 μm. N Quantification of senescence cells green area signal at different time points after co-culture, n = 3, two-way ANOVA . PBS, phosphate-buffered saline. WT, wild type. KO, knocked out. Data were represented as the mean ± SD.* p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The C57BL/6J mice and SD rats were obtained from the Animal Center of Army Medical University, NLRP3 flox/flox mice and Lyz2-Cre mice were constructed by Cyagen (GuangZhou, China) and used to generate macrophage-specific Nlrp3 conditional knockout mice (NLRP3 Lyz−/− ).

Techniques: In Situ, Western Blot, Quantitative RT-PCR, Gene Expression, Comparison, Co-Culture Assay, Cell Culture, Expressing, Flow Cytometry, Saline

Journal: Journal of Translational Medicine

Article Title: Targeting the NLRP3 in macrophages contributes to senescence cell clearance in radiation-induced skin injury

doi: 10.1186/s12967-025-06204-z

Figure Lengend Snippet: Knockout of Nlrp3 in vivo result in the accumulation of senescence cells. A Schematic diagram of the workflow for establishing a RISI skin damage model in WT mouse and Nlrp3 Lyz−/− mouse. B Representative macroscopic images of WT mouse and Nlrp3 Lyz−/− mouse at different times after radiation. C Scores of WT mouse and Nlrp3 Lyz−/− mouse at different times after radiation, n = 5, two-way ANOVA . D Representative HE staining images of WT mouse and Nlrp3 Lyz−/− mouse at different times after radiation. Scale bar, 100 μm. E Representative immunofluorescence staining images and statistics of F4/80 at different time points after radiation in WT mouse and Nlrp3 Lyz−/− mouse, n = 5, t test. Scale bar, 100 μm. F , G Representative western blot and statistics of senescence-related proteins P16, P21, P53, γ-H2AX, LMNB1 in WT mouse and Nlrp3 Lyz−/− mouse, t test. H – J Representative immunofluorescence staining images and statistics of senescence-related proteins γ-H2AX, P21, P16 in WT mouse and Nlrp3 Lyz−/− mouse at different time points after radiation, n = 5, t test. Scale bar, 100 μm. K RT-qPCR analysis of cell senescence-related genes Cdkn1a , Cdkn2a , and Lmnb1 in WT mouse and Nlrp3 Lyz−/− mouse at different time points after radiation, n = 3. L , M Heatmap and volcano plot of differentially expressed genes in the RNA sequencing of WT mouse and Nlrp3 Lyz−/− mouse 20 days after radiation. N GO analysis of differentially expressed genes in WT mouse and Nlrp3 Lyz−/− mouse 20 days after radiation. FC, fold change. Data were represented as the mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The C57BL/6J mice and SD rats were obtained from the Animal Center of Army Medical University, NLRP3 flox/flox mice and Lyz2-Cre mice were constructed by Cyagen (GuangZhou, China) and used to generate macrophage-specific Nlrp3 conditional knockout mice (NLRP3 Lyz−/− ).

Techniques: Knock-Out, In Vivo, Staining, Immunofluorescence, Western Blot, Quantitative RT-PCR, RNA Sequencing

Journal: Journal of Translational Medicine

Article Title: Targeting the NLRP3 in macrophages contributes to senescence cell clearance in radiation-induced skin injury

doi: 10.1186/s12967-025-06204-z

Figure Lengend Snippet: IL-33 secreted by senescence cells leads to the disorder of macrophage NLRP3 activation. A RT-qPCR analysis of Il33 gene at different time points after the primary dermal fibroblasts receiving X-ray induced senescence, n = 3, one-way ANOVA with Tukey HSD post hoc comparison. B , C Flow cytometry analysis and statistics of IL-33 protein expression after 7 days of primary dermal fibroblasts treated with X-ray, n = 3, t test. D , E Representative images and histograms of immunofluorescence staining for IL-33 before and after senescence induction in MSF, L929, C166, and HACAT cells. Scale bar, 50 μm. F RT-qPCR analysis of Nlrp3 and Il1b gene expression levels in iBMDM cells treated with LPS and different concentrations of IL-33, n = 3, one-way ANOVA with Tukey HSD post hoc comparison. G Western blot analysis of NLRP3 protein expression levels in iBMDM cells treated with LPS and different concentrations of IL-33. H Schematic diagram of subcutaneous injection of IgG or IL-33 neutralizing antibodies after exposure of mouse to ionizing radiation. I Western blot was used to assess the expression levels of NLRP3, CASPASE1, and IL-18 in tissues after treatment with IgG or IL-33 neutralizing antibodies. J The expression levels of senescence-related proteins P16, P21 and P53 in tissues after treatment with IgG or IL-33 neutralizing antibodies were assessed using western blot. K Representative images and quantitative analysis of P16 immunofluorescence staining after IgG or IL-33 neutralizing antibody treatment, n = 5, one-way ANOVA with Tukey HSD post hoc comparison. Scale bar, 100 μm. i.d., intradermal injection. MFI, mean fluorescence intensity. LPS, lipopolysaccharide. Data were represented as the mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The C57BL/6J mice and SD rats were obtained from the Animal Center of Army Medical University, NLRP3 flox/flox mice and Lyz2-Cre mice were constructed by Cyagen (GuangZhou, China) and used to generate macrophage-specific Nlrp3 conditional knockout mice (NLRP3 Lyz−/− ).

Techniques: Activation Assay, Quantitative RT-PCR, Comparison, Flow Cytometry, Expressing, Immunofluorescence, Staining, Gene Expression, Western Blot, Injection, Fluorescence

Journal: Journal of Translational Medicine

Article Title: Targeting the NLRP3 in macrophages contributes to senescence cell clearance in radiation-induced skin injury

doi: 10.1186/s12967-025-06204-z

Figure Lengend Snippet: Nr-CWS reduce the load of senescence cells and alleviate RISI. A Schematic diagram of the workflow for treating RISI rats with gel dressings loaded with Nr-CWS or PBS. B , C Representative macroscopic images and scoring curves of rats receiving each treatment at various time points after RISI, n = 5, one-way ANOVA with Tukey HSD post hoc comparison. D Representative hematoxylin and eosin (H&E) staining images of skin tissue at various time points in rats receiving different treatments. Scale bar, 100 μm. E ELISA assessment of plasma IL1α concentration in rats following different treatments, n = 3, one-way ANOVA with Tukey HSD post hoc comparison. F Representative immunofluorescence staining images and statistics of NLRP3 in skin tissue from rats following different treatments 10 days after radiation, n = 3, one-way ANOVA with Tukey HSD post hoc comparison, Scale bar, 100 μm. G Representative images and statistics of immunofluorescence staining of P21 in skin tissue from rats that received different treatments 10 days after radiation, n = 3, one-way ANOVA with Tukey HSD post hoc comparison, Scale bar, 100 μm. H Representative images and statistics of phagocytosis evaluation after PBS or Nr-CWS treatment of iBMDM, n = 3, t test. Scale bar, 50 μm. I Representative images and statistics of Transwell analysis after PBS or Nr-CWS treatment of iBMDM, n = 3, t test. Scale bar 50 μm. Nr-CWS, CWS, Nocardia rubra cell wall skeleton. iBMDM, immortalized bone marrow-derived macrophages. IR, ionizing radiation. ELISA, enzyme-linked immunosorbent assay. Data were represented as the mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The C57BL/6J mice and SD rats were obtained from the Animal Center of Army Medical University, NLRP3 flox/flox mice and Lyz2-Cre mice were constructed by Cyagen (GuangZhou, China) and used to generate macrophage-specific Nlrp3 conditional knockout mice (NLRP3 Lyz−/− ).

Techniques: Comparison, Staining, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Concentration Assay, Immunofluorescence, Derivative Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Helicobacter pylori infection promotes M1 macrophage polarization and gastric inflammation by activation of NLRP3 inflammasome via TNF/TNFR1 axis

doi: 10.1186/s12964-024-02017-7

Figure Lengend Snippet: NLRP3 is mainly expressed in myeloid cells, and is significantly increased in H. pylori -positive gastritis than in H. pylori -negative gastritis. A ScRNA-seq analysis of human gastritis, intestinal metaplasia, and gastric cancer tissues was carried out in our previous studies, with raw sequencing data made available in the GEO database (GSE249874). The UMAP plot (left panel) and violin plot (right panel) showing the expression of NLRP3 in different cell populations. B The UMAP plots showing the expression of NLRP3 in H. pylori -positive and -negative gastritis tissues, respectively. C Immunofluorescence staining for CD68 (green) and NLRP3 (red) in human gastritis specimens with or without H. pylori infection. Scale bar, 10 μm. D Western blotting analysis for NLRP3 expression in H. pylori -infected and uninfected gastritis tissues. E , F Immunohistochemistry staining for NLRP3 expression in human gastritis tissues with or without H. pylori infection ( n = 15 for each group). Representative images ( E ) and histological scores ( F ) were shown respectively. Scale bar, 10 μm. *, P < 0.05; ** , P < 0.01; ***, P < 0.001 . Data are expressed as the means ± SDs

Article Snippet: The male transgenic hypergastrinemic insulin-gastrin (INS-GAS) mice (#018149), aged 6–8 weeks old and on a FVB/N background, and Nlrp3 -knockout (KO) mice (#021302) were purchased from The Jackson Laboratory and housed under a 12-h light/dark cycle with ad libitum access to food and water.

Techniques: Sequencing, Expressing, Immunofluorescence, Staining, Infection, Western Blot, Immunohistochemistry

Journal: Cell Communication and Signaling : CCS

Article Title: Helicobacter pylori infection promotes M1 macrophage polarization and gastric inflammation by activation of NLRP3 inflammasome via TNF/TNFR1 axis

doi: 10.1186/s12964-024-02017-7

Figure Lengend Snippet: H. pylori infection promotes NLRP3 inflammasome activation in macrophages. A , B ELISA for human IL-1β secretion in the supernatant of PMA-induced differentiated THP1 cells following infection with H. pylori PMSS1 strain at different MOI ( A ) or different time points ( B ). Data are expressed as the means ± SDs. C ELISA for IL-18 concentration in THP1 cells cocultured with H. pylori PMSS1 or NCTC11637 strain. D , E Western blots for NLRP3 inflammasome at different MOI after co-culture of THP1 cells with H. pylori PMSS1 ( D ) or NCTC11637 ( E ) strain. F Western blots for NLRP3 inflammasome-related proteins in THP1 cells infected with PMSS1 or NCTC11637 strain at indicated time points. G Immunofluorescence staining for NLRP3 (green) in THP1 cells following H. pylori infection for 24 h. Scale bar, 10 μm. H Representative immunofluorescence images (left panel) and quantification (right panel) of ASC speck formation after stimulation of THP1 cells with H. pylori . Scale bar, 10 μm. I Representative images (upper panel) and quantitative densitometric analysis (lower panel) of western blots for NLRP3 inflammasome proteins in THP1 cells infected with 26,695 and its VacA knockout mutant strain for 24 h. *, P < 0.05; ** , P < 0.01; ***, P < 0.001

Article Snippet: The male transgenic hypergastrinemic insulin-gastrin (INS-GAS) mice (#018149), aged 6–8 weeks old and on a FVB/N background, and Nlrp3 -knockout (KO) mice (#021302) were purchased from The Jackson Laboratory and housed under a 12-h light/dark cycle with ad libitum access to food and water.

Techniques: Infection, Activation Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Western Blot, Co-Culture Assay, Immunofluorescence, Staining, Knock-Out, Mutagenesis

Journal: Cell Communication and Signaling : CCS

Article Title: Helicobacter pylori infection promotes M1 macrophage polarization and gastric inflammation by activation of NLRP3 inflammasome via TNF/TNFR1 axis

doi: 10.1186/s12964-024-02017-7

Figure Lengend Snippet: H. pylori infection causes progressive gastritis and NLRP3 inflammasome activation in the gastric tissues of INS-GAS mice. A H&E and AB-PAS staining showing the histopathological features in gastric tissues of H. pylori -infected INS-GAS mice at 4 months post-infection. Scale bar, 20 μm. B The gastric inflammation, atrophy and metaplasia scores of categorical lesions in H. pylori -infected INS-GAS mice. Control group, n = 6; H. pylori group, n = 9. Each symbol is a different mouse. C ELISA assay showing the serum levels of TNFα and IL1β in H. pylori -infected INS-GAS mice at 4 months. D Western blots showing the expression of NLRP3 inflammasome-related proteins in gastric tissues of INS-GAS mice with or without H. pylori infection. Densitometric analysis of the immunoblots is depicted below. E Immunofluorescence staining for colocalization of CD68 and NLRP3 in gastric tissues of H. pylori -infected INS-GAS mice. Scale bar, 10 μm. The western blot results were representative of three independent experiments. The results were expressed as mean ± SEM of at least three independent experiments. * P < 0.05 , **P < 0.01 , *** P < 0.001 , NS, not significant

Article Snippet: The male transgenic hypergastrinemic insulin-gastrin (INS-GAS) mice (#018149), aged 6–8 weeks old and on a FVB/N background, and Nlrp3 -knockout (KO) mice (#021302) were purchased from The Jackson Laboratory and housed under a 12-h light/dark cycle with ad libitum access to food and water.

Techniques: Infection, Activation Assay, Staining, Control, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Immunofluorescence

Journal: Cell Communication and Signaling : CCS

Article Title: Helicobacter pylori infection promotes M1 macrophage polarization and gastric inflammation by activation of NLRP3 inflammasome via TNF/TNFR1 axis

doi: 10.1186/s12964-024-02017-7

Figure Lengend Snippet: NLRP3 is necessary for H. pylori -induced M1 macrophage polarization through the STAT3 and NF-κB pathways. A WT and Nlrp3 -KO BMDMs were infected with H. pylori PMSS1 for 6 h. Western blot was performed to detect NLRP3 inflammasome proteins, such as NLRP3, Pro-IL-1β, and cleaved IL-1β, to confirm the efficiency of NLRP3 knockout. B IL1β concentration was determined by ELISA assay using the collected supernatant from the BMDMs treated in A. C WT and Nlrp3 -KO BMDMs were infected with PMSS1, and the expression of M1 macrophage signature genes was measured by qRT-PCR. D & E THP1 cells were transfected with NLRP3 siRNA and then co-cultured with the PMSS1 strain. Western blotting was performed to detect the NLRP3 inflammasome protein levels ( D ). THP1 cell supernatant was collected for detection of IL-1β concentration by ELISA ( E ). F After the knockdown of NLRP3 and H. pylori PMSS1 strain treatment, M1 macrophage signature genes were measured by qRT-PCR. G and H WT and Nlrp3 -KO BMDMs were infected with the PMSS1 strain for 6 h ( G ) and 1 h ( H ), respectively. Western blotting was performed to detect the proteins of STAT3 ( G ) and NF-κB pathways ( H ), respectively. I and J THP1 cells were transfected with NLRP3 siRNA and then infected with PMSS1 strain for 6 h ( I ) or 1 h ( J ). Western blotting analysis was performed to study the proteins of STAT3 ( I ) and NF-κB pathways ( J ), respectively. * P < 0.05 , **P < 0.01 , *** P < 0.001

Article Snippet: The male transgenic hypergastrinemic insulin-gastrin (INS-GAS) mice (#018149), aged 6–8 weeks old and on a FVB/N background, and Nlrp3 -knockout (KO) mice (#021302) were purchased from The Jackson Laboratory and housed under a 12-h light/dark cycle with ad libitum access to food and water.

Techniques: Infection, Western Blot, Knock-Out, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Transfection, Cell Culture, Knockdown

Journal: Cell Communication and Signaling : CCS

Article Title: Helicobacter pylori infection promotes M1 macrophage polarization and gastric inflammation by activation of NLRP3 inflammasome via TNF/TNFR1 axis

doi: 10.1186/s12964-024-02017-7

Figure Lengend Snippet: TNFα stimulates NLRP3 inflammasome activation in macrophages. A KEGG pathway enrichment analysis was performed using the upregulated genes in NLRP3 high macrophages, in comparison with the NLRP3 low macrophages, according to our previous scRNA data of human gastric tissues. The dotted red box marked the enriched TNF signaling pathway. B Western blots showing the levels of NLRP3 inflammasome-related proteins in THP1 cells treated with different concentrations of TNF-α (0, 100, 200, and 400 ng/ml). C Western blots displaying the expression of NLRP3 inflammasome proteins in THP1 cells treated with TNF-α (200 ng/ml) in combination with TNFα inhibitor QNZ (400 ng/ml). D Western blots presenting the levels of NLRP3 inflammasome-related proteins in THP1 cells treated with TNF-α (200 ng/ml) in combination with TNFR1 antagonist Atrosab (1 μM). E and F qRT-PCR analysis of marker genes for M1 macrophages, including TNF-α , IL-1β , IL-6 , CCL2 , and CCL3 , in THP1 cells, following stimulation with TNF and TNFα inhibitor QNZ ( E ), or with TNF and TNFR1 antagonist Atrosab ( F ). * P < 0.05 , **P < 0.01 , *** P < 0.001

Article Snippet: The male transgenic hypergastrinemic insulin-gastrin (INS-GAS) mice (#018149), aged 6–8 weeks old and on a FVB/N background, and Nlrp3 -knockout (KO) mice (#021302) were purchased from The Jackson Laboratory and housed under a 12-h light/dark cycle with ad libitum access to food and water.

Techniques: Activation Assay, Comparison, Western Blot, Expressing, Quantitative RT-PCR, Marker

Journal: Cell Communication and Signaling : CCS

Article Title: Helicobacter pylori infection promotes M1 macrophage polarization and gastric inflammation by activation of NLRP3 inflammasome via TNF/TNFR1 axis

doi: 10.1186/s12964-024-02017-7

Figure Lengend Snippet: H. pylori induces NLRP3 inflammasome and promotes M1 macrophage polarization through TNFα. A The UMAP plots indicated TNF expression in human gastric tissues from our scRNA-seq data. B Immunofluorescence staining for the co-expression of CD68 (green) and TNFα (red) in human gastritis tissues with or without H. pylori infection. Scale bar, 10 μm. C Western blot assay for TNFα protein expression in THP1 cells infected with H. pylori at various MOIs for 24 h (upper panel), or treated with H. pylori at an MOI of 100 for indicated time points (lower panel). D ELISA assay for detection of the TNF-α concentration in the supernatant of THP1 cells following H. pylori infection with indicated MOI (left panel) or at indicated time points (right panel). E Western blots for the TNFα expression in THP1 cells infected with H. pylori 26,695 strain or its VacA − mutant. F and G Western blots for the protein expression of NLRP3 inflammasome in THP1 cells treated with TNF-α in combination with TNFα inhibitor QNZ ( F ), or in combination with TNFR1 antagonist Atrosab ( G ). H and I The qRT-PCR analysis of mRNA levels of M1 macrophage signature genes in THP1 cells treated with TNF-α in combination with QNZ ( H ), or in combination with Atrosab ( I ). * P < 0.05 , **P < 0.01

Article Snippet: The male transgenic hypergastrinemic insulin-gastrin (INS-GAS) mice (#018149), aged 6–8 weeks old and on a FVB/N background, and Nlrp3 -knockout (KO) mice (#021302) were purchased from The Jackson Laboratory and housed under a 12-h light/dark cycle with ad libitum access to food and water.

Techniques: Expressing, Immunofluorescence, Staining, Infection, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay, Mutagenesis, Quantitative RT-PCR

Journal: Cell Communication and Signaling : CCS

Article Title: Helicobacter pylori infection promotes M1 macrophage polarization and gastric inflammation by activation of NLRP3 inflammasome via TNF/TNFR1 axis

doi: 10.1186/s12964-024-02017-7

Figure Lengend Snippet: TNF/TNFR1 axis is responsible for H. pylori -induced NLRP3 inflammasome activation and M1 macrophage activation. A , B CellChat analysis was used to identify cell–cell communication signaling network between H. pylori -positive and H. pylori -negative gastritis tissues. CellChat calculates communication probabilities at the signaling pathway by summarizing all ligand-receptor interactions associated with each signaling pathway. If the ratio of the total pathway probabilities between the comparison group and the control group is < 0.95 and p -value < 0.05, the communication strength in the control group was considered as significant (y-axis in red); whereas if the total ratio is > 1.05 and p -value < 0.05, the communication strength in the comparison group was considered as significant (y-axis in blue). H. pylori -positive tissues as the comparison group; H. pylori -negative tissues as control group. A The bar charts showed the most significant differential signaling pathways. The dotted red box highlighted TNF signaling as the top upregulated pathway in H. pylori -positive gastritis tissues, compared to H. pylori -negative tissues. B The heatmap plot showing differential cellular communication networks among different cell populations in H. pylori -positive and -negative gastritis tissues. The TNF signaling network was highlighted with a red box. C WT and Tnfr1 -KO BMDMs were infected with H. pylori PMSS1 strain (MOI = 100). Western blots for the protein expression of molecules of NLRP3 inflammasome. D ELISA for IL-1β concentration in cell supernatant. E The qRT-PCR analysis for M1 macrophage signature genes expression in TNFα or PMSS1-treated BMDMs from WT and Tnfr1 -deficient mice. F The working model of the TNF/TNFR1 axis-mediated activation of NLRP3 inflammasome in macrophages and M1 polarization during H. pylori infection. H. pylori infection induces the production of TNFα, which binds to TNFR1, promoting the NLRP3 inflammasome activation in macrophages, and resulting in M1 macrophage polarization and gastric inflammation through activation of NF-κB and STAT3 signaling pathways

Article Snippet: The male transgenic hypergastrinemic insulin-gastrin (INS-GAS) mice (#018149), aged 6–8 weeks old and on a FVB/N background, and Nlrp3 -knockout (KO) mice (#021302) were purchased from The Jackson Laboratory and housed under a 12-h light/dark cycle with ad libitum access to food and water.

Techniques: Activation Assay, Comparison, Control, Protein-Protein interactions, Infection, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Glyburide Suppresses Inflammation-Related Colorectal Tumorigenesis Through Inhibition of NLRP3 Inflammasome

doi: 10.3390/ijms252111640

Figure Lengend Snippet: Effects of glyburide or NLRP3 deficiency on colitis induced by DSS (experiment 1). ( A ) Chemical structure of glyburide. ( B ) Study protocol of DSS-induced colitis. ( C ) Body weight change in the experimental mice. Percentage of weight loss at the end of the study compared to the beginning. ( D ) Colon length measured from the ileocecal junction to the anal verge. ( E ) Representative photographs of colon sections stained with hematoxylin and eosin. Enlarged pictures of the section enclosed within the square in the upper panels are shown in the corresponding lower panels. Scale bars: 100 µm for upper panels and 50 µm for lower panels. ( F ) Inflammation scores. Data represent mean ± standard error. * p < 0.05. CTRL, control (C57BL/6J) mice; DSS, dextran sodium sulfate; GLB, glyburide; KO, knockout.

Article Snippet: Male C57BL/6J mice and NLRP3-KO mice with a C57BL/6J background, B6.129S6- Nlrp3 tm1Bhk /J (Strain 021302), aged 10 weeks, were obtained from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Staining, Control, Knock-Out

Journal: International Journal of Molecular Sciences

Article Title: Glyburide Suppresses Inflammation-Related Colorectal Tumorigenesis Through Inhibition of NLRP3 Inflammasome

doi: 10.3390/ijms252111640

Figure Lengend Snippet: Effect of GLB and NLRP3 deficiency on expression levels of inflammatory cytokines in the colon of a mouse colitis model. mRNA expression levels of Il1b , Il6 , and Tnf in the colonic mucosa were measured by qRT-PCR with specific primers. Data represent mean ± standard error. * p < 0.05. CTRL, control C57BL/6J mice; DSS, dextran sodium sulfate; GLB, glyburide; KO, knockout.

Article Snippet: Male C57BL/6J mice and NLRP3-KO mice with a C57BL/6J background, B6.129S6- Nlrp3 tm1Bhk /J (Strain 021302), aged 10 weeks, were obtained from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Expressing, Quantitative RT-PCR, Control, Knock-Out

Journal: International Journal of Molecular Sciences

Article Title: Glyburide Suppresses Inflammation-Related Colorectal Tumorigenesis Through Inhibition of NLRP3 Inflammasome

doi: 10.3390/ijms252111640

Figure Lengend Snippet: Effects of glyburide or NLRP3 deficiency on inflammation-related colon tumorigenesis induced by AOM/DSS (experiment 2). ( A ) Study protocol of a mouse AOM/DSS-induced CRC model. ( B ) Body weight change in the experimental mice. Percentage of weight gain at the end of the study compared to the beginning. ( C ) Blood glucose levels at the end of the experiment. ( D ) Representative macroscopic pictures of tumors in the colorectum of the experimental mice. ( E ) Representative pathological image of the colon tumor in each experimental group. Scale bars: 500 µm. ( F ) Number of tumors per mouse colon. Data represent mean ± standard error. * p < 0.05. AOM, azoxymethane; CTRL, control C57BL/6J mice; DSS, dextran sodium sulfate; GLB, glyburide; KO, knockout.

Article Snippet: Male C57BL/6J mice and NLRP3-KO mice with a C57BL/6J background, B6.129S6- Nlrp3 tm1Bhk /J (Strain 021302), aged 10 weeks, were obtained from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Control, Knock-Out

Journal: International Journal of Molecular Sciences

Article Title: Glyburide Suppresses Inflammation-Related Colorectal Tumorigenesis Through Inhibition of NLRP3 Inflammasome

doi: 10.3390/ijms252111640

Figure Lengend Snippet: Effects of GLB and NLRP3 deficiency on gene expressions in the colon of a mouse colon tumorigenesis model. mRNA expression levels of ( A ) Il1b , Il6 , and Tnf , and of ( B ) Cyclind1 , Ptgs2 , and Tgfb in the colonic mucosa were measured by qRT-PCR with specific primers. Data represent mean ± standard error. * p < 0.05. AOM, azoxymethane; CTRL, control C57BL/6J mice; DSS, dextran sodium sulfate; GLB, glyburide; KO, knockout.

Article Snippet: Male C57BL/6J mice and NLRP3-KO mice with a C57BL/6J background, B6.129S6- Nlrp3 tm1Bhk /J (Strain 021302), aged 10 weeks, were obtained from Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Expressing, Quantitative RT-PCR, Control, Knock-Out

Journal: Molecular Medicine Reports

Article Title: Hyperglycemia induces microglial pyroptosis by increasing oxygen extraction rate: Implication in neurological impairment during ischemic stroke

doi: 10.3892/mmr.2024.13270

Figure Lengend Snippet: HG treatment aggravated neurological damage after IR. (A) Blood glucose levels of the mice at 0 h after ischemia. (B) Blood glucose levels of the mice at 2 h after ischemia. (C) Blood glucose levels of the mice at 24 h after ischemia. (D) TTC staining images showed cerebral infarction (black arrow) in the IR group. The incidence of cerebral infarction was 100% in the IR group, while in the Sham group it was 0% (P<0.01). (E) Latency to fall of the Sham, IR, IR + HG, IR + NG and IR + HG + NLRP3-/- groups. (F) Foot fault of the Sham, IR, IR + HG, IR + NG and IR + HG + NLRP3-/- groups. Data are presented as the mean ± standard deviation. *P<0.05; **P<0.01; ns, not significant. n=6 mice per group. IR, ischemia-reperfusion; NG, normal glucose; HG, high glucose; NLRP3, NOD-like receptor protein 3.

Article Snippet: The NLRP3 knockout (KO) C57BL/6 mice (NLRP3 −/− ; male; age, 6 weeks) were purchased from GemPharmatech Co., Ltd.

Techniques: Staining, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Hyperglycemia induces microglial pyroptosis by increasing oxygen extraction rate: Implication in neurological impairment during ischemic stroke

doi: 10.3892/mmr.2024.13270

Figure Lengend Snippet: NLRP3 −/− inhibited microglial pyroptosis activation after IR. (A) Western blot analysis of NLRP3 (110 kDa), caspase-1 (10 kDa), GSDMD-FL (53 kDa), GSDMD-N (30 kDa), IL-1β (17 kDa) and IL-18 (22 kDa). (B) NLRP3 expression. (C) Caspase-1 expression. (D) GSDMD-FL expression. (E) GSDMD-N expression. (F) IL-1β expression. (G) IL-18 expression. Data are presented as the mean ± standard deviation. *P<0.05; **P<0.01; ns, not significant. n=6 mice per group. IR, ischemia-reperfusion; NLRP3, NOD-like receptor protein 3; GSDMD-FL, full-length gasdermin D; GSDMD-N, gasdermin D-N domain; NLRP3 −/− ; NLRP3 knockout; WT, wild-type.

Article Snippet: The NLRP3 knockout (KO) C57BL/6 mice (NLRP3 −/− ; male; age, 6 weeks) were purchased from GemPharmatech Co., Ltd.

Techniques: Activation Assay, Western Blot, Expressing, Standard Deviation, Knock-Out

Journal: Molecular Medicine Reports

Article Title: Hyperglycemia induces microglial pyroptosis by increasing oxygen extraction rate: Implication in neurological impairment during ischemic stroke

doi: 10.3892/mmr.2024.13270

Figure Lengend Snippet: NLRP3 −/− inhibited HG-induced exacerbation of pyroptosis after IR in vivo . (A) Western blot analysis of Caspase-1 (10 kDa), GSDMD-FL (53 kDa), GSDMD-N (30 kDa), IL-1β (17 kDa) and IL-18 (22 kDa). (B) Caspase-1 expression. (C) GSDMD-FL expression. (D) GSDMD-N expression. (E) IL-1β expression. (F) IL-18 expression. Data are presented as the mean ± standard deviation. *P<0.05; **P<0.01; ns, not significant. n=6 mice per group. IR, ischemia-reperfusion; NG, normal glucose; HG, high glucose; NLRP3, NOD-like receptor protein 3; GSDMD-FL, full-length gasdermin D; GSDMD-N, gasdermin D-N domain; NLRP3 −/− ; NLRP3 knockout; WT, wild-type.

Article Snippet: The NLRP3 knockout (KO) C57BL/6 mice (NLRP3 −/− ; male; age, 6 weeks) were purchased from GemPharmatech Co., Ltd.

Techniques: In Vivo, Western Blot, Expressing, Standard Deviation, Knock-Out

Journal: Molecular Medicine Reports

Article Title: Hyperglycemia induces microglial pyroptosis by increasing oxygen extraction rate: Implication in neurological impairment during ischemic stroke

doi: 10.3892/mmr.2024.13270

Figure Lengend Snippet: NLRP3 −/− inhibited HG-induced exacerbation of microglial pyroptosis after IR in vivo . (A) Immunofluorescence images showing the expression of Iba1 + microglial cells (green), Caspase-1 (red), and the co-localization of Caspase-1 and microglial cells. (B) Immunofluorescence images showing the expression of Iba1+ microglial cells (green), GSDMD-N (red), and the co-localization of GSDMD-N and microglial cells. (C) Immunofluorescence images showing the expression of Iba1+ microglial cells (green), IL-1β (red), and the co-localization of IL-1β and microglial cells. (D) Immunofluorescence images showing the expression of Iba1+ microglial cells (green), IL-18 (red), and the co-localization of IL-18 and microglial cells. Scale bars (a-i): 20 µm. (E) The fluorescence density of Caspase-1. (F) The fluorescence density of GSDMD-N. (G) The fluorescence density of IL-1β. (H) The fluorescence density of IL-18. Data are presented as the mean ± standard deviation. *P<0.05; **P<0.01; ns, not significant. n=6 mice per group. IR, ischemia-reperfusion; NG, normal glucose; HG, high glucose; NLRP3, NOD-like receptor protein 3; GSDMD-N, gasdermin D-N domain; NLRP3 −/− ; NLRP3 knockout; WT, wild-type.

Article Snippet: The NLRP3 knockout (KO) C57BL/6 mice (NLRP3 −/− ; male; age, 6 weeks) were purchased from GemPharmatech Co., Ltd.

Techniques: In Vivo, Immunofluorescence, Expressing, Fluorescence, Standard Deviation, Knock-Out

Journal: Molecular Medicine Reports

Article Title: Hyperglycemia induces microglial pyroptosis by increasing oxygen extraction rate: Implication in neurological impairment during ischemic stroke

doi: 10.3892/mmr.2024.13270

Figure Lengend Snippet: NLRP3 −/− and inhibition of Caspase-1 reversed HG-induced exacerbation of pyroptosis after OGD/IR. (A) Western blot analysis of Caspase-1 (10 kDa) in vivo . (B) Caspase-1 expression levels in vivo . (C) Caspase-1 expression levels in vitro . (D) Western blot analysis of Caspase-1 (10 kDa) in vitro . (E) GSDMD-FL expression. (F) GSDMD-N expression. (G) Western blot analysis of GSDMD-FL (53 kDa), GSDMD-N (30 kDa), IL-1β (17 kDa) and IL-18 (22 kDa). (H) IL-1β expression. (I) IL-18 expression. Data are presented as the mean ± standard deviation. *P<0.05; **P<0.01; ns, not significant. n=4 per group in vitro , n=6 per group in vivo . IR, ischemia-reperfusion; OGD, oxygen and glucose deprivation; NG, normal glucose; HG, high glucose; NLRP3, NOD-like receptor protein 3; GSDMD-FL, full-length gasdermin D; GSDMD-N, gasdermin D-N domain; NLRP3 −/− ; NLRP3 knockout; WT, wild-type; CON, control.

Article Snippet: The NLRP3 knockout (KO) C57BL/6 mice (NLRP3 −/− ; male; age, 6 weeks) were purchased from GemPharmatech Co., Ltd.

Techniques: Inhibition, Western Blot, In Vivo, Expressing, In Vitro, Standard Deviation, Knock-Out, Control

Journal: Molecular Medicine Reports

Article Title: Hyperglycemia induces microglial pyroptosis by increasing oxygen extraction rate: Implication in neurological impairment during ischemic stroke

doi: 10.3892/mmr.2024.13270

Figure Lengend Snippet: NLRP3 −/− and inhibition of Caspase-1 reversed HG-induced exacerbation of pyroptosis after OGD/IR. (A) Immunofluorescence images showing the expression of Iba1 + microglial cells (green), Caspase-1 (red), and the co-localization of caspase-1 and microglial cells. (B) Immunofluorescence images showing the expression of Iba1 + microglial cells (green), GSDMD-N (red), and the co-localization of GSDMD-N and microglial cells. (C) Immunofluorescence images showing the expression of Iba1+ microglial cells (green), IL-1β (red), and the co-localization of IL-1β and microglial cells. (D) Immunofluorescence images showing the expression of Iba1+ microglial cells (green), IL-18 (red), and the co-localization of IL-18 and microglial cells. Scale bars (a-i): 20 µm. (E) The fluorescence density of Caspase-1. (F) The fluorescence density of GSDMD-N. (G) The fluorescence density of IL-1β. (H) The fluorescence density of IL-18. Data are presented as the mean ± standard deviation. **P<0.01; ns, not significant. n=4 per group. OGD, oxygen and glucose deprivation; NG, normal glucose; HG, high glucose; NLRP3, NOD-like receptor protein 3; GSDMD-N, gasdermin D-N domain; NLRP3 −/− ; NLRP3 knockout; WT, wild-type; CON, control.

Article Snippet: The NLRP3 knockout (KO) C57BL/6 mice (NLRP3 −/− ; male; age, 6 weeks) were purchased from GemPharmatech Co., Ltd.

Techniques: Inhibition, Immunofluorescence, Expressing, Fluorescence, Standard Deviation, Knock-Out, Control