Journal: Journal of Molecular Neuroscience

Article Title: VX-765 Alleviates β-Amyloid Deposition and Secondary Degeneration in the Ipsilateral Hippocampus and Ameliorates Cognitive Decline after Focal Cortical Infarction in Rats

doi: 10.1007/s12031-022-02088-6

Figure Lengend Snippet: VX-765 improved cognitive decline after ischemia, as revealed by the Morris water maze test. a The latency of the rats to reach the platform was measured to assess the memory ability of the three groups after a 5-day training trial following ischemia. b No significant difference was observed in swimming speed among the 3 groups. c The percentage of time spent in the target quadrant for the 3 groups. d The numbers of platform crossings recorded in the 3 groups. e The swimming paths of the sham, vehicle, and VX-765 groups on the last day of the test. n = 12 per group. Data are shown as the mean ± SEM. * P < 0.05 and ** P < 0.01 versus sham group; # P < 0.05 versus vehicle group

Article Snippet: At 1 h after dMCAO, the caspase-1 inhibitor VX-765 (50 mg/kg, MedChem Express, USA) was intraperitoneally injected once daily for 28 consecutive days after dMCAO as previously described (Flores et al. ).

Techniques:

Journal: Journal of Molecular Neuroscience

Article Title: VX-765 Alleviates β-Amyloid Deposition and Secondary Degeneration in the Ipsilateral Hippocampus and Ameliorates Cognitive Decline after Focal Cortical Infarction in Rats

doi: 10.1007/s12031-022-02088-6

Figure Lengend Snippet: VX-765 attenuates secondary degeneration in the ipsilateral hippocampus at 28 days following dMCAO. a Nissl images showing intact neurons and immunofluorescent images of b NeuN, c GFAP, and d Iba-1 in the ipsilateral hippocampus after dMCAO. Scale bar: 100 μm. e Immunoblotting analysis of the expression of NeuN, GFAP, and Iba-1 in the ipsilateral hippocampus after dMCAO, n = 6 per group. Data are shown as the mean ± SEM. ** P < 0.01 versus the sham group; # P < 0.05 and ## P < 0.01 versus the vehicle group

Article Snippet: At 1 h after dMCAO, the caspase-1 inhibitor VX-765 (50 mg/kg, MedChem Express, USA) was intraperitoneally injected once daily for 28 consecutive days after dMCAO as previously described (Flores et al. ).

Techniques: Western Blot, Expressing

Journal: Journal of Molecular Neuroscience

Article Title: VX-765 Alleviates β-Amyloid Deposition and Secondary Degeneration in the Ipsilateral Hippocampus and Ameliorates Cognitive Decline after Focal Cortical Infarction in Rats

doi: 10.1007/s12031-022-02088-6

Figure Lengend Snippet: VX-765 decreases Aβ deposition and synaptic damage in the ipsilateral hippocampus 28 days following dMCAO. a Immunostaining showing Aβ accumulation in the ipsilateral hippocampus after dMCAO. Scale bar: 100 μm. b Immunoblotting images displaying BDNF, PSD95, Syna, and GAP43 levels in the ipsilateral hippocampus after dMCAO, n = 6 per group. Data are shown as the mean ± SEM. ** P < 0.01 versus the sham group; # P < 0.05 and ## P < 0.01 versus the vehicle group

Article Snippet: At 1 h after dMCAO, the caspase-1 inhibitor VX-765 (50 mg/kg, MedChem Express, USA) was intraperitoneally injected once daily for 28 consecutive days after dMCAO as previously described (Flores et al. ).

Techniques: Immunostaining, Western Blot

Journal: Journal of Molecular Neuroscience

Article Title: VX-765 Alleviates β-Amyloid Deposition and Secondary Degeneration in the Ipsilateral Hippocampus and Ameliorates Cognitive Decline after Focal Cortical Infarction in Rats

doi: 10.1007/s12031-022-02088-6

Figure Lengend Snippet: VX-765 inhibits pyroptosis in the ipsilateral hippocampus 28 days after dMCAO. a Representative immunofluorescent staining of caspase-1 in the ipsilateral hippocampus after dMCAO. Scale bar: 100 μm. b Western blot images indicating the expression of caspase-1, NLRP3, ASC, GSDMD, IL-1β, and IL-8 in the ipsilateral hippocampus after dMCAO, n = 6 per group. Data are shown as the mean ± SEM. ** P < 0.01 versus the sham group; # P < 0.05 versus the vehicle group

Article Snippet: At 1 h after dMCAO, the caspase-1 inhibitor VX-765 (50 mg/kg, MedChem Express, USA) was intraperitoneally injected once daily for 28 consecutive days after dMCAO as previously described (Flores et al. ).

Techniques: Staining, Western Blot, Expressing

Journal: Journal of Molecular Neuroscience

Article Title: VX-765 Alleviates β-Amyloid Deposition and Secondary Degeneration in the Ipsilateral Hippocampus and Ameliorates Cognitive Decline after Focal Cortical Infarction in Rats

doi: 10.1007/s12031-022-02088-6

Figure Lengend Snippet: VX-765 reduces TUNEL positivity and apoptosis in the ipsilateral hippocampus 28 days following dMCAO. a TUNEL cells in the ipsilateral hippocampus following dMCAO. Scale bar: 100 μm. b Western blot images showing the expression of Bcl-2, Bcl-xl, Bax, and cC3 in the ipsilateral hippocampus following dMCAO, n = 6 per group. Data are shown as the mean ± SEM. ** P < 0.01 versus the sham group; # P < 0.05 and ## P < 0.01 versus the vehicle group

Article Snippet: At 1 h after dMCAO, the caspase-1 inhibitor VX-765 (50 mg/kg, MedChem Express, USA) was intraperitoneally injected once daily for 28 consecutive days after dMCAO as previously described (Flores et al. ).

Techniques: TUNEL Assay, Western Blot, Expressing

Journal: Journal of Molecular Neuroscience

Article Title: VX-765 Alleviates β-Amyloid Deposition and Secondary Degeneration in the Ipsilateral Hippocampus and Ameliorates Cognitive Decline after Focal Cortical Infarction in Rats

doi: 10.1007/s12031-022-02088-6

Figure Lengend Snippet: VX-765 attenuates secondary degeneration by inhibiting the NF-κB and MAPK pathways after dMCAO. a Western blot images indicating the expression of p-NF-κB and p-IκBα in the ipsilateral hippocampus after dMCAO. b Western blot images showing the expression of p–c-Jun, p-JNK, and p-p38 in the ipsilateral hippocampus after dMCAO, n = 6 per group. Data are shown as the mean ± SEM. ** P < 0.01 versus the sham group; # P < 0.05 and ## P < 0.01 versus the vehicle group

Article Snippet: At 1 h after dMCAO, the caspase-1 inhibitor VX-765 (50 mg/kg, MedChem Express, USA) was intraperitoneally injected once daily for 28 consecutive days after dMCAO as previously described (Flores et al. ).

Techniques: Western Blot, Expressing

Journal: Cell Death Discovery

Article Title: Osteocytic Lipocalin-2 regulates bone formation locally through iron-dependent ferroptosis and Wnt suppression

doi: 10.1038/s41420-026-02956-9

Figure Lengend Snippet: qPCR analysis of Lcn2 and osteoblast and osteocyte markers ( Phex, Runx2, Dmp1, Fgf23, Sost ) during osteogenic differentiation of A BMSCs and B OCY454 osteocyte-like cells. p < 0.05 vs. week 0, n = 4 biological replicates per time point. C – I OCY454 cells were treated with recombinant murine LCN2 (rmLCN2, 100 ng/mL) ± the ferroptosis inhibitor Deferoxamine (DFO, 100 µM) for 24 h. C Quantification of FerroOrange staining showing elevated Fe²⁺ accumulation; D , E total and Fe³⁺ iron concentrations; F , G DCFDA fluorescence showing increased ROS; H , I C11-BODIPY fluorescence indicating lipid peroxidation. DFO co-treatment rescued all parameters. n = 3–4 biological replicates per group. J Annexin V/PI flow-cytometric analysis showing increased cell death with LCN2, fully rescued by DFO but not by inhibitors of apoptosis (DEVD), necroptosis (Necrostatin-1), or pyroptosis (VX-765), confirming ferroptosis as the predominant mode of cell death. K – P Loss-of-function validation in OCY454 cells transfected with Lcn2 shRNA or scrambled control. K , L , M qPCR and Western blot confirming knockdown efficiency; N , O , P flow-cytometric analysis showing reduced ROS (DCFDA, N), lipid peroxidation (C11-BODIPY, O), and cell death (Annexin V/PI, P) following Lcn2 silencing. n = 3–4 biological replicates per group. Data are presented as mean ± SD. * p < 0.05 vs. control or scrambled shRNA; $ p < 0.05 vs. LCN2 alone; one-way ANOVA with Newman-Keuls post hoc test or Student’s t test as appropriate. Data in ( A – P ) are representative of three independent experiments.

Article Snippet: 16–18 h post-transfection, cells were treated with rmLCN2 (L) or control (C) for an additional 24 h. For iron overload, OCY454 cells were cultured in the presence of ferric ammonium chloride (FAC, 250 μM, Sigma; Cat# 158040) for 24 h, while rmLCN2 treatment was reduced to 6 h. To assess cell death pathways, cells were treated with rmLCN2 (100 ng/mL) in the presence of selective inhibitors: the ferroptosis inhibitor deferoxamine (DFO, 100 μM, Tocris Cat# 14595), the necroptosis inhibitor necrostatin-1 (Nec-1, 20 μM, Tocris Cat# 11658), the apoptosis inhibitor DEVD (10 μM, Tocris Cat# 14414), and the pyroptosis inhibitor VX-765 (10 μM, Tocris Cat#28825).

Techniques: Recombinant, Staining, Fluorescence, Biomarker Discovery, Transfection, shRNA, Control, Western Blot, Knockdown

Journal: Cell Death & Disease

Article Title: A mechanism for increased sensitivity of acute myeloid leukemia to mitotoxic drugs

doi: 10.1038/s41419-019-1851-3

Figure Lengend Snippet: Survival rates of AML cells and healthy PBMCs treated with mitoxantrone (a) , doxorubicin (b) , CCCP (c) , or 2-DG (d) at LD50 doses. For each drug, cells were also tested with or without the following cell death pathway inhibitors: z-VAD-fmk (40 μM, pan-caspase inhibitor), 3-methyladenine (5 mM, autophagy inhibitor), or VX-765 (10 μM, caspase-1 inhibitor). Results show the mean ± SD from at least three replicates. Statistical testing was performed by Student’s t -test with independent samples. *** p < 0.001; ** p < 0.01; * p < 0.05; ns: p > 0.05

Article Snippet: Stock solutions of carbonyl cyanide m -chloropheny lhydrazone (CCCP, Sigma-Aldrich), mitoxantrone (MTX, Biotang, Lexington, MA, USA), doxorubicin (DOX, Ark Pharm Inc., Arlington Heights, IL, USA), cytarabine (ara-C, Accela ChemBio Ink, San Diego, CA, USA), lonidamine (Tocris, Minneapolis, MN, USA), pan-caspase inhibitor z-VAD-fmk (Apexbio Technology, Houston, TX, USA), caspase-1 inhibitor VX-765 (Apexbio Technology), and ABT-199 (ThermoFisher, Waltham, MA, USA) were dissolved in DMSO, aliquoted, and stored at −20 °C.

Techniques:

Journal: Frontiers in Immunology

Article Title: Mitochondrial damage and IL-1β production in monocytes caused by Neospora caninum infection are mediated by dense granule protein 7 and prohibitins

doi: 10.3389/fimmu.2025.1408992

Figure Lengend Snippet: IL-1β and TNF-α production in THP-1 cells and inflammasome-reconstructed 293T cells. (A) THP-1 cells were infected with the parental strain Nc1 or the NcGRA6-, NcGRA7-, or NcGRA14-deficient (KO) parasites at a multiplicity of infection (MOI) of 2.5 or treated with medium only (mock). At 20 h postinfection, the culture supernatants were collected for analysis. (B) 293T cells were transfected with inflammasome-reconstruction plasmids encoding NLRP3, ASC, procaspase-1, and pro-IL-1β, together with NcGRA7 cDNA or an empty vector. Untransfected 293T cells were used as a negative control (no plasmid). At 20 h posttransfection, the culture supernatants were collected for analysis. (C–H) THP-1 cells were pretreated with 10 μM MCC950 (an NLARP3 inhibitor), 100 μM VX765 (a CASP1 inhibitor), and 18 μM SN50 (an NF-κB inhibitor) for 2 hr and then infected with the Nc1 strain of N. caninum at a MOI of 2.5 or treated with medium only (mock). At 20 h postinfection, the culture supernatants were collected for analysis. Each value represents the mean ± SD of 4 replicates (technical replicates) in one representative experiment. Each experiment (biological replicate) was repeated two (G, H) , three (A, D, E, F) and four times (B, C) . Statistically significant differences according to one-way ANOVA or two-way ANOVA and a Tukey–Kramer post hoc analysis (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: MCC950 (an inhibitor of NLRP3 inflammasome activation by the inhibition of IL-1β release; Cayman Chemical), VX765 (an inhibitor of NLRP3 inflammasome activation by the inhibition of caspase 1 and IL-1β cleavage and release; LKT Laboratories), SN50 (an inhibitor of the nuclear transcription factor κB; Selleck, Houston, TX, USA), a carbonyl cyanide m-chlorophenyl hydrazone (CCCP, a uncoupling agent for oxidative phosphorylation that inhibits mitochondrial function; FUJFILM Wako Pure Chemical Corporation), rocaglamid A (Cayman, Ann Arbor, MI, USA) with biochemical research grades were used in this study.

Techniques: Infection, Transfection, Plasmid Preparation, Negative Control