phycoerythrin pe cy7 conjugated anti cd11b  (Thermo Fisher)

Journal: Frontiers in Cellular Neuroscience

Article Title: The Time Course of Monocytes Infiltration After Acoustic Overstimulation

doi: 10.3389/fncel.2022.844480

Figure Lengend Snippet: Inflammatory monocytes from the bloodstream infiltrated the cochlea after acoustic overstimulation. (A) Effects of acoustic overstimulation (120 dB, 1 h white band noise exposure) on the permanent threshold shift ( N = 8 ears). (B) Effects of noise exposure on outer hair cell degeneration ( N = 4 ears in each group). (C) Effects of acoustic stimulation on peripheral blood CD11b + Ly6G – F4/80 + CX3CR1 + populations and distinction between Ly6C ++ inflammatory monocytes and Ly6C – patrolling monocytes. (D) Effects of acoustic overstimulation on cell populations (CD11b + Ly6G – F4/80 + CX3CR1 + cells; Ly6C ++ possible inflammatory monocytes and Ly6C – possible resident macrophages) in cochlea at 2 days. (E) Clodronate liposome successfully reduced monocytes in peripheral blood (left) compared to neutrophils (right) in 1 dpn at which the monocytes begin to infiltrate to the cochlea. (F) Effects of clodronate liposome treatment after acoustic overstimulation on Ly6C ++ possible inflammatory monocyte populations in the cochlea after 2 days. (left) Effects of clodronate liposome treatment after acoustic overstimulation on Ly6C – possible resident macrophage populations. (right) Clod : clodronate liposome-treated group. ** : p < 0.01, *: p < 0.05, ns : not significant. (G) Immunofluorescence study comparing control and clodronate liposome at 2 dpn. White arrows indicate clusters of monocytes. The black box in the light microscopy image located on the left indicates the anatomical location of the immunofluorescence images. Scale bar = 50 μm.

Article Snippet: 7-Aminoactinomycin D (7-AAD; cat. no. 420403; Biolegend) was used for live cell gating, phycoerythrin (PE)-Cy7-conjugated anti-CD11b (M1/70; cat. no. 25-0112-82; Invitrogen, Carlsbad, CA, United States) was used for myeloid cell gating, PE-conjugated anti-Ly6G (1A8; cat. no. 127608; Biolegend) was used for neutrophil gating, APC-conjugated anti-F4/80 (BM8; cat. no. 17-4801-82; Invitrogen) was used for monocyte/macrophage gating, and APC-Cy7-conjugated anti-Ly6C (HK1.4; cat. no. 128026; Biolegend) was used for inflammatory monocyte gating ( , ).

Techniques: Immunofluorescence, Light Microscopy

Journal: Frontiers in Cellular Neuroscience

Article Title: The Time Course of Monocytes Infiltration After Acoustic Overstimulation

doi: 10.3389/fncel.2022.844480

Figure Lengend Snippet: Inflammatory monocytes infiltrated within 2 days after acoustic stimulation, whereas neutrophil infiltration was not observed. (A) Percentages of cells sorted by Ly6C expression from the CD11b + Ly6G – F4/80 + CX3CR1 + population, demonstrating Ly6C ++ cell infiltration within 2 days after acoustic overstimulation. (B) Total cell counts showing the CD11b + Ly6G – F4/80 + CX3CR1 + population did not change after 2 dpn. (C) Distinction between Ly6G + CX3CR1 – neutrophils and CD11b + cells in the peripheral blood. (D) Lack of distinct neutrophils in the cochlea at 1 day after acoustic overstimulation compared with CD11b + cells. (E) Changes in the percentages and the cell count of neutrophils in cochlea after acoustic overstimulation. There was no statistically significant value. Ctr : control, N1d : 1 day after noise, N2d : 2 days after noise, N3d : 3 days after noise, N5d : 5 days after noise, Mo : monocytes, M φ: macrophages. ****: p < 0.0001, ***: p < 0.001, **: p < 0.01, *: p < 0.05, when compared with the control. (F) Immunofluorescence study of lower portion of spiral ligament at 1 dpn. Neutrophil (white) is mixed in the cluster of inflammatory monocytes (red and weak green). Scale bar = 20 μm.

Article Snippet: 7-Aminoactinomycin D (7-AAD; cat. no. 420403; Biolegend) was used for live cell gating, phycoerythrin (PE)-Cy7-conjugated anti-CD11b (M1/70; cat. no. 25-0112-82; Invitrogen, Carlsbad, CA, United States) was used for myeloid cell gating, PE-conjugated anti-Ly6G (1A8; cat. no. 127608; Biolegend) was used for neutrophil gating, APC-conjugated anti-F4/80 (BM8; cat. no. 17-4801-82; Invitrogen) was used for monocyte/macrophage gating, and APC-Cy7-conjugated anti-Ly6C (HK1.4; cat. no. 128026; Biolegend) was used for inflammatory monocyte gating ( , ).

Techniques: Expressing, Cell Counting, Immunofluorescence

Journal: Frontiers in Cellular Neuroscience

Article Title: The Time Course of Monocytes Infiltration After Acoustic Overstimulation

doi: 10.3389/fncel.2022.844480

Figure Lengend Snippet: Infiltrated monocytes transformed into macrophages within 5 days after acoustic stimulation. Concatenated contour plots (merging four biological replicates for each time point) showing CD11b + Ly6G – F4/80 + CX3CR1 + cells at different time points after acoustic overstimulation. Black contour : cochlea sample, red contour : control peripheral blood sample, dots : center of each population.

Article Snippet: 7-Aminoactinomycin D (7-AAD; cat. no. 420403; Biolegend) was used for live cell gating, phycoerythrin (PE)-Cy7-conjugated anti-CD11b (M1/70; cat. no. 25-0112-82; Invitrogen, Carlsbad, CA, United States) was used for myeloid cell gating, PE-conjugated anti-Ly6G (1A8; cat. no. 127608; Biolegend) was used for neutrophil gating, APC-conjugated anti-F4/80 (BM8; cat. no. 17-4801-82; Invitrogen) was used for monocyte/macrophage gating, and APC-Cy7-conjugated anti-Ly6C (HK1.4; cat. no. 128026; Biolegend) was used for inflammatory monocyte gating ( , ).

Techniques: Transformation Assay

Journal: PLoS Pathogens

Article Title: Kupffer Cells Hasten Resolution of Liver Immunopathology in Mouse Models of Viral Hepatitis

doi: 10.1371/journal.ppat.1002061

Figure Lengend Snippet: (A,B) Relative frequency (A) and absolute number (B) of Gr-1 high CD11b + neutrophils (PMNs) recovered from the livers of mice that received the indicated treatment, 2 days (A) or at the indicated time point (B) after intravenous injection of 10 7 HBV-specific CTL. (C) Representative immunohistochemical micrographs of control (left) or Clo-L-treated (right) HBV transgenic livers, two days after intravenous injection of 10 7 HBV-specific CTL. HMGB-1 staining in brown. Note the nuclear-to-cytoplasm translocation of HMGB-1 in hepatocytes surrounded by PMNs (arrowhead). Scale bar represents 150 µm. n = 3 (D and E) Mean sALT activity (units/liter) measured at the indicated time points after intravenous injection of 10 7 HBV-specific CTL in HBV transgenic mice that received the indicated treatment. Irr, irrelevant antibodies. n = 6. All data are expressed as mean ± standard deviation and are representative of at least 3 independent experiments that gave similar results; differences between CTL-injected mice treated or not with Clo-L (A,B) or between Clo-L- and CTL-injected mice treated or not with αGr-1 antibodies (E) were not statistically significant unless otherwise indicated, * p <0.05, ** p <0.001.*

Article Snippet: Cells were surface-stained with phycoerythrin (PE)-conjugated anti-CD4 (clone RM4-5; BD Pharmingen) and anti-CD11c (clone HL3; BD Pharmingen); Pacific Blue-conjugated anti-CD8 (clone 53-6.7; BD Pharmingen) and anti-CD3 (clone 145-2c11; BD Pharmingen); PE-Cy7-conjugated anti-CD11b (clone M1/70; BD Pharmingen); allophycocyanin (APC)-conjugated anti-TCR (clone H57-597; BD Pharmingen) and anti-Ly6G (clone 1A8; BD Pharmingen); fluorescein isothiocyanate (FITC)-conjugated anti-Gr-1 (clone RB6-8C5; BD Pharmingen).

Techniques: Injection, Immunohistochemical staining, Transgenic Assay, Staining, Translocation Assay, Activity Assay, Standard Deviation