Journal: International Journal of Molecular Medicine

Article Title: Hypothermic machine perfusion protects DCD graft liver from ischemia-reperfusion injury by enhancing macrophage efferocytosis via KLF2-NLRP3 signaling

doi: 10.3892/ijmm.2026.5756

Figure Lengend Snippet: KLF2 inhibits the NLRP3-mediated pyroptosis pathway during CS/Rep in ECs. (A) Heatmap and (B) volcano plot of differentially expressed genes between control and ov-KLF2 HUVECs after CS/Rep treatment. mRNA levels of (C) KLF2 and (D) NLRP3 in ov-control and ov-KLF2 group of HUVECs following CS/Rep treatment were quantified using reverse transcription-quantitative PCR. (E) Co-IP analysis of the interaction between KLF2 and NLRP3. (F) Microstructure of HUVECS. (G) Representative Western blot images for KLF2, NLRP3, GSDMD, Caspase-1 and IL-18 in CS/Rep HUVECs. Quantitative analysis of KLF2 (H), NLRP3 (I), GSDMD (J), Caspase-1 (K) and IL-18 (L) protein expression based on western blot results from HUVECS of ov-control and ov-KLF2 groups. Quantitative analysis of KLF2 (M), NLRP3 (N), GSDMD (O), Caspase-1 (P) and IL-18 (Q) protein expression based on western blot results from HUVECS of sh-control and sh-KLF2 groups. (n=3). * P<0.05, ** P<0.01, *** P<0.001. KLF2, Kruppel-like Factor 2; CS/Rep, cold storage/reperfusion; HUVEC, human umbilical vein endothelial cells; ov, overexpression; IP, immunoprecipitation; GSDMD, gasdermin D; sh, short hairpin;; FC, fold-change.

Article Snippet: The incubation period between transfection and subsequent treatment was 72 h. To verify whether NLRP3 inhibition rescues the phenotypes resulting from KLF2 deficiency, sh-KLF2-transfected cells were treated (37°C) with 10 μ M NLRP3 inhibitor MCC950 (cat. no. HY12815, MedChemExpress) for 24 h and subjected to CS/Rep as aforementioned.

Techniques: Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Co-Immunoprecipitation Assay, Western Blot, Expressing, Over Expression, Immunoprecipitation

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Experimental design. A total of 82 mice (male 42 and female 40) of both HD and WT were treated with MCC950 solution or vehicle (drinking water) from 6 to 20 weeks of age. Depression and anxiety‐like behaviour was evaluated at week 10 and 11. Cognitive function was assessed from 12 to 13 weeks of age. Motor function was evaluated every week by clasping score and rotarpd performance. At week 14, gait was assessed to further evaluate the motor function by digigait analysis. Gut function was evaluated at week 15 and 20 by GTT and faecal output and water content measurement. At week 20, gut permeability and gut macroscopy were assessed. Abbreviations: GTT, gut transit time; HD, Huntington's disease; MWM, Morris water maze; NOR, novel object recognition; NSFT, novelty‐suppressed feeding test; WT, wild type. Figure created in Biorender.com .

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Permeability

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Effects of MCC950 on phenotypic features of HD and WT littermate control mice. Body weight (a, b), food intake (c, d), fluid intake (e, f), brain weight (g, h). Data are displayed as mean ± SEM. Statistical analyses were performed using linear mixed model repeated measure analysis/generalised LMM repeated measure analysis (a–f) and linear mixed model (LMM)/generalised LMM (g–h) followed by Bonferroni post hoc adjustment with emmeans package in R . p value ( α ) = 0.05. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05. n = 10–11 (male), n = 10 (female). Symbols in Figure , indicate the significance of the post hoc comparisons as follows: *HD (H2O vs MCC), #WT (H2O vs MCC), @H2O (WT vs HD), $MCC (WT vs HD) and for each comparsion one to four symbols in the graph represent p < 0.05 to p < 0.0001, respectively. HD, Huntington's disease; MCC, MCC950; WT, wild type.

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Control

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Effects of MCC950 on motor function of HD and WT littermate control. Clasping score (a, b), rotarod performance (c, d), digigait (e, f). Data are displayed as mean ± SEM. Statistical analyses were performed using CLMMs (cumulative link mixed models) (a‐b), linear mixed model repeated measure analysis/generalised LMM repeated measure analysis (c, d), and linear mixed model (LMM)/generalised LMM (e, f) followed by Bonferroni post hoc adjustment with emmeans package in R. p value ( α ) = 0.05 and **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, ns= non‐significant n = 10–11 (male), n = 10 (female) for clasping score and rotarod; n = 8–10 (male), n = 9–10 (female) for digigait. Symbols in Figure , indicate the significance of post hoc comparisons as follows: *H2O (HD vs WT), #WT (H2O vs MCC950), $HD (H2O vs MCC950). Also, in Figure and , # symbol represents the significant interaction between treatment and age. One to four symbols in the graph represent p < 0.05 to p < 0.0001, respectively. HD, Huntington's disease; WT, wild type.

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Control

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Effects of MCC950 on short‐term memory and learning of HD and WT littermate control mice. Novel object recognition (NOR) test (a–c), Y‐maze test (d–g). Data are displayed as mean ± SEM. Statistical analyses were performed using linear mixed model (LMM)/generalised LMM (a–c), generalised linear model (d) and linear mixed model/generalised LMM repeated measure analysis (c, f, g) followed by Bonferroni post hoc adjustment with emmeans package in R. p value ( α ) = 0.05 and **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, ns = non‐significant. n = 10–11 (male), n = 10 (female). HD, Huntington's disease; WT, wild type.

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Control

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Effects of MCC950 on longer‐term associative cognition and spatial memory of HD and WT littermate control mice (CFC test). Fear conditioning (a, b) and fear extinction (c, d). Data are displayed as mean ± SEM. Statistical analyses were performed using linear mixed model repeated measure analysis/generalised LMM repeated measure analysis (a–d) followed by Bonferroni post hoc adjustment with emmeans package in R. p ‐value ( α ) = 0.05 and ####,**** p < 0.0001, ###,*** p < 0.001, ##,** p < 0.01, #,* p < 0.05. n = 10–11 (male), n = 10 (female). HD, Huntington's disease; MCC, MCC950; WT, wild type.

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Control

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Effects of MCC950 on longer‐term associative cognition and spatial memory of HD and WT littermate control mice (MWM). Latency to reach the platform during training period of MWM test (a, b), distance travelled to reach the platform during training period of MWM test (c, d), time spent in each quadrant on day 6 of MWM test (e, f), latency to reach the platform on day 6 of MWM test (g), times spent in target quadrant on day 6 of MWM test (h), total distance travelled on day 6 of MWM test (i), velocity on day 6 of MWM test (j). Data are displayed as mean ± SEM. Statistical analyses were performed using linear mixed model repeated measure analysis/generalised LMM repeated measure analysis (a–f) and linear mixed model (LMM)/generalised LMM (g–j) followed by Bonferroni post hoc adjustment with emmeans package in R. p value ( α ) = 0.05 and ####,**** p < 0.0001, ###,*** p < 0.001, ##,** p < 0.01, #,* p < 0.05, ns= non‐significant. n = 8–11 (male), n = 7–10 (female). HD, Huntington's disease; MWM, Morris water maze; NE, northeast; NW, northwest; SE, southeast (target quadrant); SW, southwest; WT, wild type.

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Control

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Effects of MCC950 on the gut profile of HD and WT littermate control mice. Faecal water content (a, b), faecal consistency (Bristol stool score) (c, d), faecal output (e, f), gut transit time (GTT) (g, h), gut permeability (i, j). Data are displayed as mean ± SEM. Statistical analyses were performed using linear mixed model repeated measure analysis/generalised LMM repeated measure analysis (a, b, e–h), CLMMs (cumulative link mixed models) (c, d), linear mixed model (LMM)/generalised LMM (i, j) followed by Bonferroni post hoc adjustment with emmeans package in R. p ‐value ( α ) = 0.05 and ####,**** p < 0.0001, ###,*** p < 0.001, ##,** p < 0.01, #,* p < 0.05. n = 10–11 (male), n = 10 (female). HD, Huntington's disease; GTT, gut transit time; WT, wild type.

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Control, Permeability

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Effects of MCC950 on gut macroscopy of HD and WT littermate control mice. Cecal length (a, b), cecal weight (c, d), colon length (e, f), colon weight (g, h), colon weight to length ratio (i). Data are displayed as mean ± SEM. Statistical analyses were performed using linear mixed model (LMM)/generalised LMM followed by Bonferroni post hoc adjustment with emmeans package in R. p value ( α ) = 0.05 and **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, ns = non‐significant. n = 9–11 (male), n = 9–10 (female). HD, Huntington's disease; WT, wild type.

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Control

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Effect of MCC950 on gut microbiome diversity of HD and WT littermate control mice. Comparison of community (Shannon alpha diversity) of gut microbiome stratified by sex under MCC950 or water for both WT and HD mice (a). PCA plot of microbial ASV based on Aitchison distance for Beta diversity (b). n = 8 (male), n = 8 (female). Box plots show the median (central line) and interquartile range (box); whiskers represent 1.5× the interquartile range; points represent individual animals. ASV, amplicon sequence variant; HD, Huntington's disease; PCA, principal component analysis; WT, wild type.

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Control, Comparison, Amplification, Sequencing, Variant Assay

Journal: Journal of Neurochemistry

Article Title: Inhibition of the NLRP3 Inflammasome With MCC950 Improves Gut Health in Huntington's Disease Mice

doi: 10.1111/jnc.70419

Figure Lengend Snippet: Effect of MCC950 on NLRP3, IL‐1β, Caspase‐1 expression in gut (proximal colon) of HD and WT littermate control mice. NLRP3 and IL‐1β (a), caspase‐1 (b, c). Data are displayed as mean ± SEM. Statistical analyses were performed using linear model (LM)/generalised LM followed by Bonferroni post hoc adjustment with emmeans package in R. p value ( α ) = 0.05 and **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, ns = non‐significant. n = 5–7 (2 blots for males and 2 blots for females). After transferring the protein from gel to membrane, ponceau S staining was performed and then each membrane was split into two parts based on the size of the protein of interest. Upper part of the membrane was probed with NLRP3 antibody. While lower part of the membrane was first probed with IL‐1β followed by caspase‐1. Membrane was stripped using stripping buffer in between two experiments of IL‐1β and caspase‐1 detection. Representative image of ponceau S staining as loading control for both NLRP3 and IL‐1β (a), and caspase‐1 (b) is shown in NLRP3 and IL‐1β (a) panel only as it was derived from a single membrane and image was captured before probing with any antibody. Positive controls, poly I:C (spleen), BMDM: LPS + Nig. BMDM, Bone‐marrow‐derived macrophage; Cas 1 KO, caspase‐1 knock out; HD H 2 O, Huntington's disease water; HD MC, Huntington's disease MCC950, NL KO, NLRP3 knock out; IL‐1beta KO, Interleukin 1 beta knock out; IL‐1β/IL‐1beta, Interleukin 1 beta; LPS, Lipopolysaccharides; Nig, nigericin; WT H 2 O, wild type water; WT MC, wild type MCC950.

Article Snippet: Starting from 6 weeks of age and up to 20 weeks, mice were treated with MCC950 sodium (Selleck Chemicals, Catalogue No. S7809) (delivered in drinking water) versus vehicle (drinking water only) ( n = 10–11 per group).

Techniques: Expressing, Control, Transferring, Membrane, Staining, Stripping Membranes, Derivative Assay, Knock-Out