primary macrophage membrane mark antibody kit  (Cell Signaling Technology Inc)

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