Journal: Parasites & Vectors

Article Title: Preliminary evaluation of the protective effects of recombinant AMA1 and IMP1 against Eimeria stiedae infection in rabbits

doi: 10.1186/s13071-022-05492-4

Figure Lengend Snippet: Trial design and vaccine procedures

Article Snippet: The separated r Es AMA1 and r Es IMP1 were transferred onto nitrocellulose (NC) membranes (Boster, Wuhan, China), separately.

Techniques: Infection, Sterility, Injection

Journal: Parasites & Vectors

Article Title: Preliminary evaluation of the protective effects of recombinant AMA1 and IMP1 against Eimeria stiedae infection in rabbits

doi: 10.1186/s13071-022-05492-4

Figure Lengend Snippet: Multiple sequence alignments of IMP1 and AMA1 from different species. a Multiple alignment of E. stiedae IMP1 with IMP1 proteins from other parasites: Eimeria maxima (UniProt: F4MKA6), Eimeria acervulina (UniProt: U6GHD9), Eimeria tenella (UniProt: F4MKA7), Eimeria mitis (UniProt: U6JVY4), Eimeria praecox (UniProt: U6GBP2); ( b ) multiple alignment of E. stiedae AMA1 with AMA1 proteins from other parasites: Eimeria tenella (UniProt: U6KTA0), Eimeria brunetti (UniProt: U6LBB9), Toxoplasma gondii (UniProt: B6KAM0), Besnoitia besnoiti (UniProt: A0A2A9MBX4), Neospora caninum (UniProt: F0VH85); Blue shading indicates conserved residues. Dashed red boxes represent B-cell epitopes. The transmembrane region is marked with a solid red box

Article Snippet: The separated r Es AMA1 and r Es IMP1 were transferred onto nitrocellulose (NC) membranes (Boster, Wuhan, China), separately.

Techniques: Sequencing

Journal: Parasites & Vectors

Article Title: Preliminary evaluation of the protective effects of recombinant AMA1 and IMP1 against Eimeria stiedae infection in rabbits

doi: 10.1186/s13071-022-05492-4

Figure Lengend Snippet: SDS-PAGE and Western blotting analysis of r Es IMP1 ( a ) and r Es AMA1 ( b ). Lane M: Protein molecular weight markers; lanes 1–4: recombinant proteins that were dissolved in the supernatant, 4 M urea, 6 M urea, and 8 M urea after ultrasonication; lane 5: purified recombinant proteins; lane 6: purified recombinant proteins incubated with anti- E. stiedae positive sera; lane 7: purified recombinant proteins incubated with negative sera from coccidia-free rabbits (the bands are indicated using arrows)

Article Snippet: The separated r Es AMA1 and r Es IMP1 were transferred onto nitrocellulose (NC) membranes (Boster, Wuhan, China), separately.

Techniques: SDS Page, Western Blot, Molecular Weight, Recombinant, Purification, Incubation

Journal: Parasites & Vectors

Article Title: Preliminary evaluation of the protective effects of recombinant AMA1 and IMP1 against Eimeria stiedae infection in rabbits

doi: 10.1186/s13071-022-05492-4

Figure Lengend Snippet: Protective effects of r Es AMA1 and r Es IMP1 against E. stiedae infection under different evaluation indicators

Article Snippet: The separated r Es AMA1 and r Es IMP1 were transferred onto nitrocellulose (NC) membranes (Boster, Wuhan, China), separately.

Techniques: Infection

Journal: Parasites & Vectors

Article Title: Preliminary evaluation of the protective effects of recombinant AMA1 and IMP1 against Eimeria stiedae infection in rabbits

doi: 10.1186/s13071-022-05492-4

Figure Lengend Snippet: Gross postmortem examinations of the liver. a PBS-uninfected group; ( b ) PBS-infected group; ( c ) Quil-A-infected group; ( d ) Trx-His-S-infected group; ( e ) r Es AMA1 group; ( f ) r Es IMP1 group

Article Snippet: The separated r Es AMA1 and r Es IMP1 were transferred onto nitrocellulose (NC) membranes (Boster, Wuhan, China), separately.

Techniques: Infection

Journal: Parasites & Vectors

Article Title: Preliminary evaluation of the protective effects of recombinant AMA1 and IMP1 against Eimeria stiedae infection in rabbits

doi: 10.1186/s13071-022-05492-4

Figure Lengend Snippet: Biochemical estimation of ALT and AST levels in the six groups

Article Snippet: The separated r Es AMA1 and r Es IMP1 were transferred onto nitrocellulose (NC) membranes (Boster, Wuhan, China), separately.

Techniques:

Journal: Parasites & Vectors

Article Title: Preliminary evaluation of the protective effects of recombinant AMA1 and IMP1 against Eimeria stiedae infection in rabbits

doi: 10.1186/s13071-022-05492-4

Figure Lengend Snippet: Changes in serum antibody levels. The changes in serum anti-r Es IMP1 ( a ) and r Es AMA1 ( b ) antibody levels after the first vaccination (week 0, indicated by arrows), booster vaccination (week 2, indicated by arrows), and infection with E. stiedae (week 4, indicated by an arrow)

Article Snippet: The separated r Es AMA1 and r Es IMP1 were transferred onto nitrocellulose (NC) membranes (Boster, Wuhan, China), separately.

Techniques: Infection

Journal: Cell Death and Differentiation

Article Title: ZBP1 and TRIF trigger lethal necroptosis in mice lacking caspase-8 and TNFR1

doi: 10.1038/s41418-024-01286-6

Figure Lengend Snippet: a RIPK1 IHC in E11.5 Ripk1 +/+ Ripk3 −/− ( n = 1) and Ripk1 −/− Ripk3 D161N/− ( n = 1) embryos. Scale bar, 100 μm. b RIPK1 IHC (left column) and Ripk1 ISH (right column) in WT small intestines. Scale bar, 50 µm. Results representative of 5 mice. c RIPK3 IHC in lung. Note the non-specific labeling of Ripk3 −/− striated muscle and granulocytes with the Abcam polyclonal antibody, which is not seen with the Genentech 1G6 antibody. Scale bar, 100 µm. Results representative of 5 WT and 3 Ripk3 −/− mice. d Mlkl ISH in small intestine with RNAscope probes targeting the full transcript (left) and a Basescope probe (center and right) that targets exon 3, which is deleted in Mlkl −/− mice. Scale bar, 25 µm. Results representative of 5 WT and 3 Mlkl −/− mice. e ZBP1 IHC in small intestine. Scale bar, 50 µm. Results representative of 5 WT and 4 Zbp1 −/− mice.

Article Snippet: GN58.3 anti-mouse ZBP1 (Genentech) was raised against ZBP1 residues A2-I153.

Techniques: Labeling, RNAscope

Journal: Cell Death and Differentiation

Article Title: ZBP1 and TRIF trigger lethal necroptosis in mice lacking caspase-8 and TNFR1

doi: 10.1038/s41418-024-01286-6

Figure Lengend Snippet: ZBP1 IHC in mouse tissues. Scale bars, 100 μm (spleen, large intestine) or 50 µm (liver, skin). Results representative of 5 WT and 4 Zbp1 −/− mice.

Article Snippet: GN58.3 anti-mouse ZBP1 (Genentech) was raised against ZBP1 residues A2-I153.

Techniques:

Journal: Cell Death and Differentiation

Article Title: ZBP1 and TRIF trigger lethal necroptosis in mice lacking caspase-8 and TNFR1

doi: 10.1038/s41418-024-01286-6

Figure Lengend Snippet: ZBP1 and TRIF contribute to embryonic lethality of Casp8 −/− Tnfr1 −/− mice.

Article Snippet: GN58.3 anti-mouse ZBP1 (Genentech) was raised against ZBP1 residues A2-I153.

Techniques:

Journal: Cell Death and Differentiation

Article Title: ZBP1 and TRIF trigger lethal necroptosis in mice lacking caspase-8 and TNFR1

doi: 10.1038/s41418-024-01286-6

Figure Lengend Snippet: E15.5 skin ( a ), liver ( b ), and placenta ( c ) sections with ZBP1 immunolabeling (brown). Scale bars, 50 μm ( a , b ) or 100 μm ( c ). Images representative of WT ( n = 5), Tnfr1 −/− ( n = 7), Casp8 −/− Tnfr1 −/− ( n = 4), Casp8 −/− Mlkl −/− ( n = 3), and Casp8 −/− Tnfr1 −/− Zbp1 −/− ( n = 7) embryos. Arrows highlight vascular labeling in Casp8 −/− Tnfr1 −/− skin. d ZBP1 IHC scores for the embryos in a – c and their placentas. Lines indicate the mean. See methods for scoring criteria. e E15.5 liver sections with immunolabeling of F4/80 (brown). Scale bars, 100 μm. Images representative of WT ( n = 5), Casp8 −/− Tnfr1 −/− ( n = 5), Casp8 −/− Tnfr1 −/− Zbp1 −/− ( n = 7), and Casp8 −/− Mlkl −/− ( n = 3) embryos. f Heatmap shows differentially expressed genes in E15.5 Casp8 −/− Tnfr1 −/− fetal livers ( n = 3) when compared with WT ( n = 3), Tnfr1 −/− ( n = 2), and Casp8 −/− Mlkl −/− ( n = 3) fetal livers.

Article Snippet: GN58.3 anti-mouse ZBP1 (Genentech) was raised against ZBP1 residues A2-I153.

Techniques: Immunolabeling, Labeling

Journal: Cell Death and Differentiation

Article Title: ZBP1 and TRIF trigger lethal necroptosis in mice lacking caspase-8 and TNFR1

doi: 10.1038/s41418-024-01286-6

Figure Lengend Snippet: a E17.5 skin sections stained with H&E or immunolabeled with phospho-RIPK3 T 231 , S 232 (pRIPK3, brown). Scale bars, 100 μm. Graphs show dermatitis histology scores and pRIPK3 IHC scores for Tnfr1 −/− Zbp1 −/− ( n = 5), Casp8 −/− Tnfr1 −/− Zbp1 −/− ( n = 4), Tnfr1 −/− Trif −/− ( n = 2), and Casp8 −/− Tnfr1 −/− Trif −/− ( n = 4) embryos. Lines indicate the mean. b Hepatocellular vacuolation histology scores for the embryos in a . Lines indicate the mean. See methods for scoring criteria used in a and b . c Representative H&E-stained skin sections from E18.5 Casp8 −/− Tnfr1 −/− Trif −/− Zbp1 −/− ( n = 3) and Tnfr1 −/− Trif −/− Zbp1 −/− ( n = 3) embryos demonstrating epidermal thickening and increased dermal cellularity in the Casp8 −/− Tnfr1 −/− Trif −/− Zbp1 −/− embryo. Scale bar, 50 μm. d Kaplan–Meier curves of mouse survival. P -value determined by 2-sided log-rank test. e Body weights of Casp8 −/− Tnfr1 −/− Trif −/− Zbp1 −/− (red bars; females, n = 17 aged 3 weeks, n = 12 aged 5 weeks; males, n = 11 aged 3 wks, n = 10 aged 5 weeks) and Tnfr1 −/− Trif −/− Zbp1 −/− (white bars; females, n = 14 aged 3 weeks, n = 11 aged 5 weeks; males, n = 13 aged 3 wks, n = 11 aged 5 weeks) mice. Bars indicate the mean ± s.e.m. P -values determined by 2-sided student t-test with Welch’s correction. f Representative liver, lung, and spleen sections from male Casp8 −/− Tnfr1 −/− Trif −/− Zbp1 −/− mice ( n = 3) with increased cellular infiltrates in the lung and liver and increased splenic hematopoiesis and male littermate controls ( Casp8 +/- Tnfr1 −/− Trif −/− , n = 2 and Casp8 +/- Tnfr1 −/− Trif +/- Zbp1 +/- , n = 1) aged 9-10 weeks. Scale bars, 100 μm (lung, spleen) or 50 μm (liver). g Peripheral blood cell counts in Casp8 −/− Tnfr1 −/− Trif −/− Zbp1 −/− ( n = 4) and Tnfr1 −/− Trif −/− Zbp1 −/− ( n = 4) mice aged 4 weeks. Bars indicate the mea n ± s.e.m. P -values are shown if P < 0.05 by 2-sided t-test with Welch’s correction. h Spleen weight as a percentage of body weight for mice aged 4-5 weeks. Red bars, Casp8 −/− Tnfr1 −/− Trif −/− Zbp1 −/− mice ( n = 4; 1 male, 3 females). White bars, littermate Tnfr1 −/− Trif −/− Zbp1 −/− ( n = 2; 1 male, 1 female) or Tnfr1 −/− Trif −/− ( n = 1, female) mice. Bars indicate the mean ± s.e.m. P -value determined by 2-sided student t-test with Welch’s correction. i Splenic leukocyte subsets for the mice in h . Bars indicate the mean ± s.e.m. P -values are shown if P < 0.05 by 2-sided t-test with Welch’s correction. j Representative spleen flow cytometry contour plots of the mice in h .

Article Snippet: GN58.3 anti-mouse ZBP1 (Genentech) was raised against ZBP1 residues A2-I153.

Techniques: Staining, Immunolabeling, IF-P, Flow Cytometry

Journal: iScience

Article Title: BAD-mediated neuronal apoptosis and neuroinflammation contribute to Alzheimer's disease pathology

doi: 10.1016/j.isci.2021.102942

Figure Lengend Snippet: BAD is involved in activation of microglial NLRP3 inflammasome via ROS-oxidative mtDNA axis (A) Detection of BAD mitochondrial translocation in plaque-associated microglia in the cortex of 6-month-old mice (left panels). Scale bar, 5 μm. The co-localization of BAD and mitochondria was analyzed by the overlap of BAD and COX-IV immunofluorescent signals (right panels). (B) Immunoblotting analysis of caspase-1 (Casp-1) cleavage in 6-month-old mouse brain extracts. The ratio of Casp-1 p20 relative intensity to β-actin was quantified (n = 3). Data were presented as mean ± SEM. ∗∗p < 0.01, ∗∗∗p < 0.001, one-way ANOVA test. (C) ASC specks were detected in plaque-associated microglia in the cortex of 6-month-old WT, 5XFAD and 5XFAD/ Bad −/− mice. Scale bar, 20 μm. The arrows indicate ASC speck-containing microglia. z stack images showed ASC speck was localized inside of microglia. The percentage of ASC speck-containing microglia was quantified (n = 3). Data were presented as mean ± SEM. ∗∗p < 0.01, one-way ANOVA test. (D) Detection of BAD mitochondrial translocation in plaque-associated microglia in the frontal cortex of patients with Alzheimer’s disease and age-matched healthy control brains (left panels). Scale bar, 5 μm. The co-localization of BAD with mitochondria was analyzed by the overlap of BAD and COX-IV immunofluorescent signals (right panels). (E) ASC specks were detected in plaque-associated microglia in the frontal cortex of patients with Alzheimer's disease and control brains. Scale bar, 20 μm. The arrows indicate ASC speck-containing microglia. The percentage of ASC speck-containing microglia was quantified (n = 5). Data were presented as mean ± SEM. ∗∗∗p < 0.001, student’s t -test. (F) Oxidized mitochondrial DNA was detected by immunofluorescent analysis with anti-8-OHdG antibody in plaque-associated microglia in the cortex of 6-month-old WT, 5XFAD and 5XFAD/ Bad −/− mice. Scale bar, 20 μm. The average area of 8-OHdG in plaque-associated microglia was quantified (n = 5). Data were presented as mean ± SEM. ∗∗p < 0.01, one-way ANOVA test. (G) Oxidized mitochondrial DNA was detected in plaque-associated microglia in the frontal cortex of patients with Alzheimer’s disease and age-matched healthy control brains. Scale bar, 20 μm. The average area of 8-OHdG in plaque-associated microglia was quantified (n = 5). Data were presented as mean ± SEM. ∗∗∗p < 0.001, student’s t -test. See also Figure S9 .

Article Snippet: Mouse anti-Caspase-1 p20 (mouse) , AdipoGen , Cat# AG-20B-0042; RRID: AB_2490248.

Techniques: Activation Assay, Translocation Assay, Western Blot, Control

Journal: iScience

Article Title: BAD-mediated neuronal apoptosis and neuroinflammation contribute to Alzheimer's disease pathology

doi: 10.1016/j.isci.2021.102942

Figure Lengend Snippet:

Article Snippet: Mouse anti-Caspase-1 p20 (mouse) , AdipoGen , Cat# AG-20B-0042; RRID: AB_2490248.

Techniques: Recombinant, Blocking Assay, Enzyme-linked Immunosorbent Assay, Isolation, Software, Microscopy, Flow Cytometry

Journal: bioRxiv

Article Title: Hypoglycemia Aggravated Cognitive Degeneration by activating Endothelial ZBP1-mediated PANoptosis in Type 2 Diabetes

doi: 10.64898/2026.03.17.712288

Figure Lengend Snippet: a , Western blotting of pyroptosis, apoptosis and necroptosis markers after knockdown ZBP1 expression in HG or HG+LG treated bEnd.3 in vitro . b , Statistical analysis of western blotting in . c, Hoechst/PI staining of bEnd.3 after ZBP1 knockdown treated with HG or HG+LG. d, Statistical analysis of PI/Hoechst ratio. e-f, Representative images and statistical analysis of ROS production in bEnd.3 after ZBP1 knockdown treated HG or HG+LG. g-h , Cell apoptosis tested by flow cytometry in bEnd.3 after knockdown ZBP1 treated with HG or HG+LG. * p <0.05 vs HG group, ** p <0.01 vs HG group, **** p <0.0001 vs HG group; # p <0.05 vs HG+LG group, ## p <0.001 vs HG+LG group, ### p <0.001 vs HG+LG group. ZBP1: Z-DNA binding protein 1.

Article Snippet: Protein samples (20 μg/lane) underwent electrophoresis in a 4%–20% SDS polyacrylamide gel (P0468S and P0469S, Beyotime) and mounted onto polyvinylidene fluoride membranes (FFP39, Beyotime) fixed with 5% milk in TBST for 1.5 h and infested with primary antibodies (NLRP3, Proteintech, 68102-1-Ig; PYRIN, Abcam, ab195975; RIPK1, Proteintech, 29932-1-AP; RIPK3, Proteintech, 17563-1-AP; Casepase-8, Proteintech, 13423-1-AP; Caspase-1, Proteintech, 22915-1-AP; ZBP1, Proteintech, 13285-1-AP;AIM2, Proteintech, 20590-1-AP; N-GSDMD, Proteintech, ab215203; GSDMD, Proteintech, 20770-1-AP; MLKL, Proteintech, 66675-1-Ig; Caspase-3, Proteintech, 66470-2-Ig; FADD, Proteintech, 14906-1-AP;ASC, Proteintech, 10500-1-AP; IL-18, Proteintech, 10663-1-AP; IL-1β, Proteintech, ab283818; Casepse-9, Proteintech, 66169-1-Ig; Bcl2, Proteintech, 12789-1-AP; BAX, Proteintech, 50599-2-Ig; β-actin, Servicebio, GB15001-100) overnight at 4°C.

Techniques: Western Blot, Knockdown, Expressing, In Vitro, Staining, Flow Cytometry, Binding Assay

Journal: bioRxiv

Article Title: Hypoglycemia Aggravated Cognitive Degeneration by activating Endothelial ZBP1-mediated PANoptosis in Type 2 Diabetes

doi: 10.64898/2026.03.17.712288

Figure Lengend Snippet: a , Volcano plot of differential expression genes in bEnd.3 after knocking down the expression of ZBP1. b , Heat map of differential expression genes in bEnd.3 after knocking down the expression of ZBP1. c , KEGG enrichment analysis bubble chart of differential genes in bEnd.3 after knocking down the expression of ZBP1. d , Immunofluorescent staining of RAGE in mouse hippocampus DG area of the DM and RH-DM group. e , Western blotting of RAGE after knocking down ZBP1 in HG or HG+LG treated bEnd.3. f , Statistical analysis of western blotting in ( n =3). g , Representative immunofluorescent staining images of RAGE in bEnd.3 after silencing ZBP1 expression in HG or HG+LG group. h , Statistical analysis of immunofluorescent density in ( n =3). i, Western blotting of RAGE and ZBP1-associated PANoptosis molecules after knocking down RAGE in HG or HG+LG treated bEnd.3. j, Statistical analysis of western blotting in ( n =3). * p <0.05 vs HG group, ** p <0.01 vs HG group, *** p <0.001 vs HG group, **** p <0.0001 vs HG group; # p <0.05 vs HG+LG group, ## p <0.01 vs HG+LG group, ### p <0.001 vs HG+LG group, #### p <0.0001 vs HG+LG group. AGEs: advanced glycation end products; RAGE: receptor for advanced glycation end products.

Article Snippet: Protein samples (20 μg/lane) underwent electrophoresis in a 4%–20% SDS polyacrylamide gel (P0468S and P0469S, Beyotime) and mounted onto polyvinylidene fluoride membranes (FFP39, Beyotime) fixed with 5% milk in TBST for 1.5 h and infested with primary antibodies (NLRP3, Proteintech, 68102-1-Ig; PYRIN, Abcam, ab195975; RIPK1, Proteintech, 29932-1-AP; RIPK3, Proteintech, 17563-1-AP; Casepase-8, Proteintech, 13423-1-AP; Caspase-1, Proteintech, 22915-1-AP; ZBP1, Proteintech, 13285-1-AP;AIM2, Proteintech, 20590-1-AP; N-GSDMD, Proteintech, ab215203; GSDMD, Proteintech, 20770-1-AP; MLKL, Proteintech, 66675-1-Ig; Caspase-3, Proteintech, 66470-2-Ig; FADD, Proteintech, 14906-1-AP;ASC, Proteintech, 10500-1-AP; IL-18, Proteintech, 10663-1-AP; IL-1β, Proteintech, ab283818; Casepse-9, Proteintech, 66169-1-Ig; Bcl2, Proteintech, 12789-1-AP; BAX, Proteintech, 50599-2-Ig; β-actin, Servicebio, GB15001-100) overnight at 4°C.

Techniques: Quantitative Proteomics, Expressing, Staining, Western Blot

Journal: bioRxiv

Article Title: Hypoglycemia Aggravated Cognitive Degeneration by activating Endothelial ZBP1-mediated PANoptosis in Type 2 Diabetes

doi: 10.64898/2026.03.17.712288

Figure Lengend Snippet: In type 2 diabetic mouse brain, when hypoglycemia happens, the sensor of PANoptosome, ZBP1, was initially activated in the endothelial cells. After that, molecules involved in pyroptosis, apoptosis and necroptosis were overtly raised in bEnd.3 cell line. Moreover, the AGEs-RAGE axis was activated by hypoglycemia and ZBP1, which aggregated the ZBP1-mediated PANoptosis in bEnd.3. Based on PANoptosis was activated, endothelial cells injured and hippocampus was in an inflammatory state thus led to spatial learning and cognitive degeneration.

Article Snippet: Protein samples (20 μg/lane) underwent electrophoresis in a 4%–20% SDS polyacrylamide gel (P0468S and P0469S, Beyotime) and mounted onto polyvinylidene fluoride membranes (FFP39, Beyotime) fixed with 5% milk in TBST for 1.5 h and infested with primary antibodies (NLRP3, Proteintech, 68102-1-Ig; PYRIN, Abcam, ab195975; RIPK1, Proteintech, 29932-1-AP; RIPK3, Proteintech, 17563-1-AP; Casepase-8, Proteintech, 13423-1-AP; Caspase-1, Proteintech, 22915-1-AP; ZBP1, Proteintech, 13285-1-AP;AIM2, Proteintech, 20590-1-AP; N-GSDMD, Proteintech, ab215203; GSDMD, Proteintech, 20770-1-AP; MLKL, Proteintech, 66675-1-Ig; Caspase-3, Proteintech, 66470-2-Ig; FADD, Proteintech, 14906-1-AP;ASC, Proteintech, 10500-1-AP; IL-18, Proteintech, 10663-1-AP; IL-1β, Proteintech, ab283818; Casepse-9, Proteintech, 66169-1-Ig; Bcl2, Proteintech, 12789-1-AP; BAX, Proteintech, 50599-2-Ig; β-actin, Servicebio, GB15001-100) overnight at 4°C.

Techniques: