Journal: Neurobiology of disease

Article Title: Age-dependent neurodegeneration and neuroinflammation in a genetic A30P/A53T double-mutated α-synuclein mouse model of Parkinson's disease.

doi: 10.1016/j.nbd.2022.105798

Figure Lengend Snippet: Fig. 9. Analysis of CD11b+ microglia in the SN and the striatum of wt and of hm2α-SYN-39 mice. (A) Exemplary images of CD11b+ microglia in the SN of 2–3 months old and 16–17 months old wt and hm2α-SYN-39 mice. High magnification inserts of single cells are included in each image. The scale bar indicates 25 μm. (B) Analysis of CD11b+ microglia in wt mice (yellow; 2–3 months: n = 8; 7–8 months: n = 8; 11–12 months: n = 7; 16–17 months: n = 7) and in hm2α-SYN-39 mice (blue; 2–3 months: n = 6; 7–8 months: n = 8; 11–12 months: n = 7; 16–17 months: n = 10) (F (7,54) = 23.86, p < .0001, eta-squared 0.76). (C) Exemplary images of CD11b+ microglia in the striatum of 2–3 months old and 16–17 months old wt and hm2α-SYN-39 mice. High magnification inserts of single cells are included in each image. The scale bar indicates 25 μm. (D) Analysis of CD11b+ microglia in wt mice (yellow; 2–3 months: n = 8; 7–8 months: n = 8; 11–12 months: n = 7; 16–17 months: n = 7) and in hm2α-SYN-39 mice (blue; 2–3 months: n = 7; 7–8 months: n = 8; 11–12 months: n = 6; 16–17 months: n = 10) (F (7,53) = 22.52, p < .0001, eta-squared 0.75). n indicates the number of biologically-independent animals, mean values for each time point ± SEM are shown. Statistical analysis was done using the one-way ANOVA followed by Tukey's post-test. *p < .05; **p < .01; ***p < .001; ****p < .0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: After fixation with 4% PFA for 15 min, the sections were incubated with rat anti-mouse CD4 (1:1000, Serotec, cat # MCA1767), rat anti-mouse CD8 (1:500, Serotec, cat # MCA609G), rat anti-mouse CD11b (1:100, Serotec, cat # MCA711), rat anti-mouse B220 (1:100, BD Pharmingen, cat # 550286) or rabbit anti-mouse TH (1:500, Abcam, cat # ab112) antibodies overnight.

Techniques:

Journal: Journal of translational medicine

Article Title: Cholesterol overload in macrophages drives metabolic dysfunction-associated steatohepatitis via inhibiting 7-dehydrocholesterol reductase in mice.

doi: 10.1186/s12967-024-05905-1

Figure Lengend Snippet: Fig. 1 Hepatic cholesterol accumulation and M1 macrophages infiltration in MASH. Six-week-old male C57BL/6J mice were fed either a NCD or a CDAHFD for six weeks (n = 3). (a) Body weight gain curves and (b) liver/body weight ratio was recorded. (c) Serum ALT and AST levels were detected. (d) Liver sections stained with H&E and ORO, and immunohistochemically for F4/80+ and CD11b+, 100× magnification. (e-f) Quantifies hepatic TC and TG levels. (g) NAS score. (h-j) Hepatic relative mRNA expression of Tnfa, Il1b and Il6. (k-l) Hepatic relative protein levels of CD86, ARG-1 and CD206. (m-o) Flow cytometry analysis of macrophages (live+CD45+F4/80+CD11b+) and M1 macrophages (live+CD45+F4/80+CD11b+iNOS+). (p-q) Flow cytometry analysis of TREM2 (live+CD45+F4/80+TREM2+). Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 between groups

Article Snippet: Immunohistochemical analysis utilized anti-F4/80 (70076s) and anti-CD11b (17800s) antibodies from Cell Signaling Technology (CST, USA), and biotinylated goat anti-rabbit IgG (SA1022, BOSTER, China).

Techniques: Staining, Expressing, Flow Cytometry

Journal: Journal of translational medicine

Article Title: Cholesterol overload in macrophages drives metabolic dysfunction-associated steatohepatitis via inhibiting 7-dehydrocholesterol reductase in mice.

doi: 10.1186/s12967-024-05905-1

Figure Lengend Snippet: Fig. 3 Correlation between DHCR7 expression and macrophage polarization in MASH. (a) Relative expression of cholesterol biosynthesis and efflux genes in primary macrophages after ox-LDL (100 µg/ mL) or ox-LDL combined with statin treating for 24 h. (b) Expression levels of cholesterol biosyn thesis and efflux genes in primary macrophages polarized to M1 with LPS (100 ng/mL) or M2 with IL-4 (20 ng/mL) for 24 h. (c-d) Western blot analysis for DHCR7 protein in M1 and M2 macrophages, with densitometry. (e) Immunofluorescence staining for DHCR7 in the polarized-macrophages, 200× magnification. (f-g) Multiple immunofluorescence co-localization for DHCR7 and CD11B on livers of NCD or CDAHFD mice. (h-i) DHCR7 protein and mRNA levels in total liver and primary macrophages isolated from livers of mice on control and CDAHFD. (j-k) Immunofluorescence multi-labeling of liver sections from NC and MASH patients for DHCR7 and macrophage marker CD68 to determine the proportion of DHCR7+CD68+ cells among total CD68+ cells. Data are presented as mean ± SD, with n = 3 per group. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 between groups

Article Snippet: Immunohistochemical analysis utilized anti-F4/80 (70076s) and anti-CD11b (17800s) antibodies from Cell Signaling Technology (CST, USA), and biotinylated goat anti-rabbit IgG (SA1022, BOSTER, China).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining, Isolation, Control, Labeling, Marker

Journal: Journal of translational medicine

Article Title: Cholesterol overload in macrophages drives metabolic dysfunction-associated steatohepatitis via inhibiting 7-dehydrocholesterol reductase in mice.

doi: 10.1186/s12967-024-05905-1

Figure Lengend Snippet: Fig. 5 Myeloid-macrophage-specific DHCR7 deficiency exacerbates MASH progression. Wildtype (WT) and myeloid-specific DHCR7 knockout (DHCR7 cKO) mice were fed with a normal control diet (CON) or a CDAHFD for 10 weeks (n = 7). (a) Body weight progression, (b) liver/body weight ratio, and (c-d) serum ALT and AST levels were detected. (e-f) Hepatic TG and TC levels were measured. (g) ELISA quantification of liver IL-1β and IL-6 levels. (h) Histological analysis of liver sections is presented through H&E, ORO staining, and immunohistochemistry for F4/80+ and CD11b+ cells (200× magnification). (i) NAS score. (j-k) Relative protein levels of CD86, ARG-1, CD206 were determined, with densitometry. (l) Flow cytometry of primary macrophage in cKO mice and WT with different diets. (m) Flow cytometry of TREM2 in macrophages. (n) Predominantly of the M1 type in macrophages, including quantification of both the percentage of M1 macrophages and M1 macrophage counts per liver weight. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 between group

Article Snippet: Immunohistochemical analysis utilized anti-F4/80 (70076s) and anti-CD11b (17800s) antibodies from Cell Signaling Technology (CST, USA), and biotinylated goat anti-rabbit IgG (SA1022, BOSTER, China).

Techniques: Knock-Out, Control, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry, Flow Cytometry

Journal: PLoS ONE

Article Title: Oxidative Stress and Proinflammatory Cytokines Contribute to Demyelination and Axonal Damage in a Cerebellar Culture Model of Neuroinflammation

doi: 10.1371/journal.pone.0054722

Figure Lengend Snippet: List of primary antibodies used for immunofluorescence studies.

Article Snippet: CD11b/OX42 , mouse anti-rat CD11b , 1∶150 , Serotec.

Techniques: Immunofluorescence, Immunopeptidomics, Purification, Binding Assay

Journal: Cancers

Article Title: Cancer Vaccination against Extracellular Vimentin Efficiently Adjuvanted with Montanide ISA 720/CpG

doi: 10.3390/cancers14112593

Figure Lengend Snippet: Immune cell infiltration in B16F10 melanoma after vaccination. ( A ) CD45+ immune cells present in B16F10 tumors derived from Study II as determined by IHC. Each dot represents one microscopic field. ( B ) Representative images of CD45 staining for each group at 200× magnification. ( C ) CD3+ immune cells present in B16F10 tumors derived from Study II as determined by IHC. Each dot represents one microscopic field. ( D ) Representative images of CD3 staining for each group at 200× magnification. ( E ) CD11b+ immune cells present in B16F10 tumors derived from Study II as determined by IHC. Each dot represents one microscopic field. ( F ) Representative images of CD11b staining for each group at 200× magnification. * p < 0.05, ** p < 0.01, and **** p < 0.0001.

Article Snippet: Next, slides were incubated with the following primary antibodies: anti-CD31 rat anti-mouse IgG2a (1:50, Dianova, Hamburg, Germany, Cat. DIA-310M, clone SZ31), anti-CD45 rabbit anti-mouse (1:100, Abcam, Cat. Ab10558), anti-CD11b rabbit anti-mouse (1:4000, Abcam, Cat. Ab13357), anti-CD3 rabbit anti-mouse (1:600, Neomarkers, Fremond, CA, USA, Cat. RM-9717-S), and mouse anti-vimentin (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. sc-373717, Clone E5, 1:50 dilution).

Techniques: Derivative Assay, Staining

Journal: Cancers

Article Title: Cancer Vaccination against Extracellular Vimentin Efficiently Adjuvanted with Montanide ISA 720/CpG

doi: 10.3390/cancers14112593

Figure Lengend Snippet: Enhanced immune cell infiltration correlates with vessel density and tumor growth ( A ) Significant correlation between the number of CD45+, CD3+, and CD11b+ immune cells and vessel density (number of CD31+ vessel per microscopic field). ( B ) Correlation between CD45+, CD3+, and CD11b+ immune cell infiltration and final tumor volume at day 14 (mm 3 ). Significant correlations ( p < 0.05) are indicated with the dark grey area.

Article Snippet: Next, slides were incubated with the following primary antibodies: anti-CD31 rat anti-mouse IgG2a (1:50, Dianova, Hamburg, Germany, Cat. DIA-310M, clone SZ31), anti-CD45 rabbit anti-mouse (1:100, Abcam, Cat. Ab10558), anti-CD11b rabbit anti-mouse (1:4000, Abcam, Cat. Ab13357), anti-CD3 rabbit anti-mouse (1:600, Neomarkers, Fremond, CA, USA, Cat. RM-9717-S), and mouse anti-vimentin (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. sc-373717, Clone E5, 1:50 dilution).

Techniques: