cd11b positive cell fraction  (Miltenyi Biotec)

Journal: Molecular Brain

Article Title: Peripheral-central network analysis of cancer cachexia status accompanied by the polarization of hypothalamic microglia with low expression of inhibitory immune checkpoint receptors

doi: 10.1186/s13041-024-01091-9

Figure Lengend Snippet: Changes in hypothalamic microglia in a mouse model of peritoneal dissemination of pancreatic cancer. A Schematic workflow for the isolation of hypothalamic microglia of pancreatic cancer cell-transplanted mice at the pre-cachexia or cachexia stage. B - G Changes in the mRNA expression of TNFα, IL-1β, IL-6, PD-1, CD112R and LCN2 in the hypothalamic CD11b + microglia of pancreatic cancer cell-transplanted mice at the pre-cachexia compared to those of control mice. Data are plotted as mean ± S.E.M. Unpaired t -test: * p < 0.05, ** p < 0.01 vs. control group (3 independent experiments). H - M Changes in the mRNA expression of TNFα, IL-1β, IL-6, PD-1, CD112R and LCN2 in the hypothalamic CD11b + microglia of pancreatic cancer cell-transplanted mice at the cachexia stage. Data are plotted as mean ± S.E.M. Unpaired t -test: * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group (6 or 7 independent experiments). N Pearson correlation scatter plot of the expression of LCN2 mRNA (ΔCt: LCN2 (Ct) -GAPDH (Ct) ) and the expression of PD-1 mRNA (ΔCt: PD-1 (Ct) -GAPDH (Ct) ) in hypothalamic microglia obtained from pancreatic cancer cell-transplanted mice at the pre-cachexia or cachexia stage. ( r = -05326, p = 0.0337) ( O ) Pearson correlation scatter plot of the expression of LCN2 mRNA (ΔCt: LCN2 (Ct) -GAPDH (Ct) ) and the expression of CD112R mRNA (ΔCt: CD112R (Ct) -GAPDH (Ct) ) in hypothalamic microglia obtained from pancreatic cancer cell-transplanted mice at the pre-cachexia or cachexia stage. ( r = -0.6704, p = 0.0045) ( P ) Increase in the number of cFos-tdTomato + cells in the PVN of pancreatic cancer cell-transplanted cfos-TRAP2-tdTomato mice at the cachexia stage (on Day 18) compared to that in control mice (Scale bar: 100 μm). Q Plasma levels of corticosterone in mice at the cachexia stage after transplantation of pancreatic cancer cells. Each value represents the mean ± S.E.M. Unpaired t -test: *** p < 0.001 vs. control group ( n = 3–4)

Article Snippet: Then, CD11b (microglia) micro beads (Miltenyi Biotec Co.) were added to the ACSA2-negative cell fraction and the CD11b + cell fraction was sorted by an auto MACS pro separator.

Techniques: Isolation, Expressing, Control, Clinical Proteomics, Transplantation Assay

Journal: Molecular Brain

Article Title: Peripheral-central network analysis of cancer cachexia status accompanied by the polarization of hypothalamic microglia with low expression of inhibitory immune checkpoint receptors

doi: 10.1186/s13041-024-01091-9

Figure Lengend Snippet: Changes in cytotoxic immune cells of the spleen and hypothalamic microglia of mice with LPS-injected systemic inflammation. A Schedule of FACS and MACS sorting after LPS injection (600 μg/0.1 mL/mouse, i.v.). B Changes in body weight at pre/post-injection of saline or LPS. Data are plotted as mean ± S.E.M. Statistical analysis was performed using two-way ANOVA with the Bonferroni test: *** p < 0.001 vs. saline-injected group ( n = 5–6). C - H Changes in the mRNA expression of TNFα, IL-1β, IL-6, PD-1, CD112R and LCN2 in the hypothalamic CD11b + microglia of LPS-injected model mice compared to those in saline-injected mice. Data are plotted as mean ± S.E.M. Unpaired t -test: * p < 0.05, ** p < 0.01 vs. saline-injected group (3 or 7 independent experiments). (I-M) Representative flow cytometric plots (left panel) and quantitative evaluation of the number (right panel) of CD8 + T cells, CD4 + T cells (I-K) and NK cells (L, M) in the spleen of LPS-injected model mice compared to those in saline-injected mice. Data are plotted as mean ± S.E.M. Unpaired t -test: * p < 0.05, ** p < 0.01, *** p < 0.001 vs. saline-injected group ( n = 4–5)

Article Snippet: Then, CD11b (microglia) micro beads (Miltenyi Biotec Co.) were added to the ACSA2-negative cell fraction and the CD11b + cell fraction was sorted by an auto MACS pro separator.

Techniques: Injection, Saline, Expressing