Journal: Veterinary Medicine and Science

Article Title: A case of generalised cutaneous apocrine cystomatosis in a Pekingese dog

doi: 10.1002/vms3.711

Figure Lengend Snippet: The antibodies and protocols used for immunohistochemical evaluation

Article Snippet: Mouse and rabbit‐specific HRP (ABC) detection IHC kit , Manufacturer instructions , , Abcam; Cambridge, UK.

Techniques: Immunohistochemical staining, Incubation

Journal: JCI insight

Article Title: CETP inhibition enhances monocyte activation and bacterial clearance and reduces streptococcus pneumonia-associated mortality in mice.

doi: 10.1172/jci.insight.173205

Figure Lengend Snippet: Figure 4. CETPi increases macrophage infiltration into site of infection and macrophage activation while maintaining tissue homeostasis at different stages of sepsis. (A and B) Representative staining figures of M1 macrophages by marker CD86 in lung obtained 0 and 72 hours after infection by S. pneu- moniae from female APOA1.CETP mice treated with control or CETPi. Quantification was determined by random sampling spots on lungs of 3 different mice in each treatment and by measuring stained areas using ImageJ (H&E 0 hours, mean ± SD, 16.0 ± 2.6 [CETPi, n = 3] versus 2.7 ± 1.5 [Control, n = 3], unpaired t test, P = 0.002) (IHC 72 hours, mean ± SD, 6.0 ± 3.5 [CETPi, n = 3] versus 17.0 ± 7.9% [Control, n = 3], unpaired t test, P = 0.02). (C and D) Propor- tion of infiltrating inflammatory macrophages (CD11b+Ly6c+) and tissue repairing and inflammation resolving macrophages (CD11b+Ly6c–) in BAL samples obtained 24 and 72 hours after infection from female APOA1.CETP mice treated with control or CETPi (CD11b+Ly6c+ 72 hours, mean ± SD, 5.6 ± 1.8 [CETPi, n = 4] versus 14.3 ± 6.1% [Control, n = 3], unpaired t test with Bonferroni correction, P = 0.037) (CD11b+ Ly6c– 24 hours, mean ± SD, 62.1 ± 6.8 [CETPi, n = 4] ver- sus 45.6 ± 3.8 [Control, n = 3], unpaired t test with Bonferroni correction, P = 0.013; CD11b+ Ly6c– 72 hours, mean ± SD, 78.3 ± 9.1 [CETPi, n = 4] versus 34.7 ± 22.7 [Control, n = 3], unpaired t test with Bonferroni correction, P = 0.016). (E) Proportion of migrating monocytes (Ly6C++Ly6GDIM) in blood samples obtained 0 and 72 hours after infection from female APOA1.CETP mice treated with control or CETPi (mean ± SD, 12.0 ± 2.7 [CETPi, n = 5] versus 0.14 ± 0.06 [Control, n = 4], unpaired t test, P = 0.00005). Data are shown as mean ± SD. *P < 0.05, **P < 0.01, ****P < 0.0001. ApoA1, apolipoprotein A-1; CETP, cholesteryl ester transfer protein; CD86, cluster of differentiation 86; CD11b, integrin α M; Ly6-C, lymphocyte antigen 6 locus C; Ly6-G, lymphocyte antigen 6 locus G.

Article Snippet: For IHC, tissue cross-sections were incubated with a primary macrophage membrane mark antibody kit (CD86 and CD206, 97624, CST) to identify specific cell types of macrophages followed by secondary antibodies using the alkaline phosphatase system to amplify signals.

Techniques: Infection, Activation Assay, Staining, Marker, Control, Sampling

Journal: JCI insight

Article Title: CETP inhibition enhances monocyte activation and bacterial clearance and reduces streptococcus pneumonia-associated mortality in mice.

doi: 10.1172/jci.insight.173205

Figure Lengend Snippet: Figure 5. CETPi decreases caspase-1 and COX-2 protein expression in human and mouse cells. (A–C) COX-2 and Caspase-1 expression in PBMCs treated with increasing doses of CETPi. (D–E) COX-2 expression in THP1 cells treated with control or CETPi (1 μM) (mean ± SD, 1.00 ± 0.42 [Control, n = 3] versus 1.31 ± 0.38 [CETPi, n = 3], unpaired t test, P = 0.03). (F–G) Caspase-1 in THP1 cells treated with control or CETPi (1 μM) (mean ± SD, 1.00 ± 0.19 [Control, n = 3] versus 2.02 ± 0.11 [CETPi, n = 3], unpaired t test, P = 0.02). (H) Expression of ROS in THP1 cells treated with control or CETPi (2 μM), (mean ± SD, 2.45 ± 0.36 [CETPi, n = 6] versus 1.00 ± 0.19 [Control, n = 6], unpaired t test, P = 0.00005). (I–K) Expression of COX-2 and CETP in RAW264.7 cells after adenovirus transfection induced-CETP overexpression in increasing MOIs. (L–M) Expression of pro–caspase-1 and cleaved caspase-1 in RAW 264.7 cells after adeno- virus transfection–induced CETP overexpression at MOI = 1.6. Pro–caspase-1: mean ± SD, 1.00 ± 0.15 (Control, n = 3) versus 0.45 ± 0.22 (CETP overexpress, n = 3), unpaired t test, P = 0.02; cleaved caspase-1: mean ± SD, 1.00 ± 0.14 (Control, n = 3) versus 0.58 ± 0.16 (CETP overexpress, n = 3), unpaired t test, P = 0.03. Data displayed as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001. Caspase-1, caspase-1/IL-1 converting enzyme; COX-2, prostaglandin-endoperoxide synthase 2; MOI, multiplicity of infection; PBMC, human peripheral blood mononuclear cell; RAW, murine macrophage cell; ROS, reactive oxygen species; THP1, human peripheral blood monocyte.

Article Snippet: For IHC, tissue cross-sections were incubated with a primary macrophage membrane mark antibody kit (CD86 and CD206, 97624, CST) to identify specific cell types of macrophages followed by secondary antibodies using the alkaline phosphatase system to amplify signals.

Techniques: Expressing, Control, Transfection, Over Expression, Virus, Infection

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: ZMAT1 acts as a tumor suppressor in pancreatic ductal adenocarcinoma by inducing SIRT3/p53 signaling pathway

doi: 10.1186/s13046-022-02310-8

Figure Lengend Snippet: Over-expression of ZMAT1 promotes apoptosis in Pancreatic Ductal Adenocarcinoma (PDAC) cells. A Flow cytometry showed that over-expression of ZMAT1 increased the percentage of apoptotic SW1990 cells. B - C Flow cytometry showed that ZMAT1 deletion reduced the percentage of apoptotic SW1990 cells. D - F Western blot showed over-expression of ZMAT1 up-regulated BAD, BAX, Cleaved Caspase 3 and declined Bcl-2 in SW1990 cells ( D ), whereas ZMAT1 knockdown reduced BAD, BAX, Cleaved Caspase 3 and increased Bcl-2 in BXPC-3 ( F ) and Capan-2 cells ( G ). All * P -value < 0.05, ** P -value < 0.01, *** P -value < 0.001. P-values were assessed using two-tailed t-tests in C-E. All figures represent mean ± SD from three independent experiments

Article Snippet: Tunel apoptosis assay kit (#8109) was from Cell Signaling Technology (Danfoss, USA).

Techniques: Over Expression, Flow Cytometry, Western Blot, Knockdown, Two Tailed Test

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: ZMAT1 acts as a tumor suppressor in pancreatic ductal adenocarcinoma by inducing SIRT3/p53 signaling pathway

doi: 10.1186/s13046-022-02310-8

Figure Lengend Snippet: ZMAT1 functions in a p53-dependent manner. A Gene set enrichment analysis (GSEA) of RNA sequencing on SW1990/Vector and SW1990/ZMAT1-OV cells groups. B Heatmap of RNA sequencing showed differential expression levels of key nodes in p53-assicoated cell cycle and apoptosis pathways. C - D RT-qPCR and western blot were used to detect the p53 levels in ZMAT1 over-expressed ( C ) and knockdown cells ( D ). E Double-label immunofluorescence staining for the intracellular localization of ZMAT1 and p53 in SW1990 cells. F luciferase activity assays were performed on 293 T cells with co-transfection of pGL3-p53 with 0.01 μg, 0.05 μg, 0.1 μg, 0.5 μg and 1 μg of vector encoding ZMAT1. G SW1990/ZMAT1-OV cells were treated with Pifithrin-α for 24 h and the protein levels of ZMAT1, p53, p21, and BAD were analyzed by immunoblotting with the indicated antibodies. H CCK-8 assays were performed on SW1990/ZMAT1-OV cells after treating with Pifithrin-α for 24 h. I Colony formation assays were performed on SW1990/ZMAT1-OV cells after treating with Pifithrin-α for 24 h. All * P -value < 0.05, ** P -value < 0.01, *** P -value < 0.001. P-values were assessed using two-tailed t-tests and ANOVA followed by Dunnett’s tests for multiple comparison in B, C, D, E, G, J and K. All figures represent mean ± SD from three independent experiments

Article Snippet: Tunel apoptosis assay kit (#8109) was from Cell Signaling Technology (Danfoss, USA).

Techniques: RNA Sequencing, Plasmid Preparation, Quantitative Proteomics, Quantitative RT-PCR, Western Blot, Knockdown, Immunofluorescence, Staining, Luciferase, Activity Assay, Cotransfection, CCK-8 Assay, Two Tailed Test, Comparison

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: ZMAT1 acts as a tumor suppressor in pancreatic ductal adenocarcinoma by inducing SIRT3/p53 signaling pathway

doi: 10.1186/s13046-022-02310-8

Figure Lengend Snippet: ZMAT1 correlates with p53 in Pancreatic Ductal Adenocarcinoma (PDAC). A BALB/c-nudes ( n = 6 per group) were sacrificed 60 days after the injection and tumors dissected from respective groups were shown. B Tumor growth curves after the injection of SW1990/Vector cells and SW1990/ZMAT1-OV cells. Tumor volume was calculated every 10 days. C Tumor weight was measured in ZMAT1-OV and control groups. D The protein levels of ZMAT1, SIRT3 and p53 of tumors were analyzed by immunoblotting with the indicated antibodies. E IHC staining of ZMAT1, SIRT3, p53, Ki67 and Tunel in tumors from ZMAT1-OV and control groups. F Representative images of double-label immunofluorescence (IF) staining of ZMAT1 and p53 in 60 PDAC tissues. G IF staining showed ZMAT1 expression level highly correlated with p53 expression. H Kaplan–Meier analysis in PDAC patients grouped according to the expression levels of ZMAT1 and p53 showed that PDAC patients with high ZMAT1/high p53 expression had the longest overall survival among all the groups. I A schematic diagram for the role of the ZMAT1-SIRT3-p53 axis in regulation of cell cycle and apoptosis in PDAC. All * P -value < 0.05, ** P -value < 0.01, *** P -value < 0.001. Scale bars: 200 μm. P -values were assessed using two-tailed t-tests and ANOVA followed by Dunnett’s tests for multiple comparison in B-C. Spearman’s correlation was performed in G. Kaplan–Meier analyses and log-rank tests were conducted in H

Article Snippet: Tunel apoptosis assay kit (#8109) was from Cell Signaling Technology (Danfoss, USA).

Techniques: Injection, Plasmid Preparation, Control, Western Blot, Immunohistochemistry, TUNEL Assay, Immunofluorescence, Staining, Expressing, Two Tailed Test, Comparison