Journal: Journal of Neuroinflammation

Article Title: Modulation of cannabinoid receptor 2 alters neuroinflammation and reduces formation of alpha-synuclein aggregates in a rat model of nigral synucleinopathy

doi: 10.1186/s12974-024-03221-5

Figure Lengend Snippet: Modulation of CB2 with SMM-189 globally reduces macrophages and infiltrating monocytes and CD4 T lymphocytes at 4 weeks. Brain immune cells from high cohort rats were isolated from lesioned (+ AAV-human Asyn) and unlesioned (-AAV-human Asyn) rat hemispheres at 4 weeks and flow cytometry analysis conducted to identify monocyte, lymphocyte and microglia populations from CD45 and CD11b expression and further gated into more distinct populations. Macrophage populations (CD163 + CD11b+) were identified from live cells (see Figure ). A ) Representative dot blots of CD163 + macrophages in the unlesioned hemisphere of a vehicle and SMM-189-treated rat at 4 weeks. B ) Quantification of CD163 + reveals a global reduction of macrophages from SMM-189 treatment. C ) Representative dot blots of CD45 and CD11b in the unlesioned hemisphere of a vehicle and SMM-189-treated rat. D ) Quantification of the monocyte population as a frequency of all live cells (CD45hiCD11b+). E ) Quantification of microglia (CD45loCD11b+), and microglia MFI of F ) CD11b and G ) MHCII. H ) Representative dot blots of CD3 + lymphocytes stratified by CD4 and CD8 populations in the lesion hemisphere of a vehicle and SMM-189-treated rat. I ) Quantification of CD4-expressing T lymphocytes. * p < 0.05

Article Snippet: In brief, endogenous peroxidase was quenched with 3% hydrogen peroxide, blocked with 8% normal serum, avidin (Vector Cat#SP-2001) and 0.1% Triton-X and incubated overnight at 4°C with biotin (Vectorlabs Cat#SP-2001), normal serum and primary antibodies specific for Asyn (Clone 4B12; BioLegend Cat# 807801, RRID: AB_2564730), pSer129 Asyn (Clone EP1536Y; Abcam Cat# ab51253, RRID: AB_869973), CD68 (Bio-Rad Cat# MCA341R, RRID: AB_2291300), MHCII (Bio-Rad Cat# MCA46GA, RRID: AB_567369), and CD163 (Bio-Rad Cat# MCA342A, RRID: AB_2074557).

Techniques: Isolation, Flow Cytometry, Expressing

Journal: Journal of Neuroinflammation

Article Title: Modulation of cannabinoid receptor 2 alters neuroinflammation and reduces formation of alpha-synuclein aggregates in a rat model of nigral synucleinopathy

doi: 10.1186/s12974-024-03221-5

Figure Lengend Snippet: Modulation of CB2 with SMM-189 promotes innate immune cell activation and altered infiltration of T lymphocytes in the brain at 8 weeks. Brain immune cells from high cohort rats were isolated from lesioned- (+ AAV-human Asyn) and unlesioned (-AAV-human Asyn) hemispheres at 8 weeks and flow cytometry analysis conducted to identify monocyte, lymphocyte and microglia populations using CD45 and CD11b expression and further gated into more distinct populations (see Figure ). A ) Representative dot blots of CD163 + macrophages in the unlesioned hemisphere of a vehicle and SMM-189-treated rat at 8 weeks. B ) Quantification of monocyte (CD45hiCD11b+) and C ) MHCII-expressing classical monocyte (MHCII + CD43-) frequencies. D ) Analysis of microglia (CD45loCD11b+) and microglia MFI of E ) MHCII F ) CD172a and G ) CD11b. H ) Dot blots of CD3 + lymphocytes stratified by CD4 and CD8 populations in the lesion hemisphere of a vehicle and SMM-189-treated rat at 8 weeks. I ) Quantification of CD4-expressing and J ) CD8-expressing T cells. * p < 0.05, ** p < 0.01

Article Snippet: In brief, endogenous peroxidase was quenched with 3% hydrogen peroxide, blocked with 8% normal serum, avidin (Vector Cat#SP-2001) and 0.1% Triton-X and incubated overnight at 4°C with biotin (Vectorlabs Cat#SP-2001), normal serum and primary antibodies specific for Asyn (Clone 4B12; BioLegend Cat# 807801, RRID: AB_2564730), pSer129 Asyn (Clone EP1536Y; Abcam Cat# ab51253, RRID: AB_869973), CD68 (Bio-Rad Cat# MCA341R, RRID: AB_2291300), MHCII (Bio-Rad Cat# MCA46GA, RRID: AB_567369), and CD163 (Bio-Rad Cat# MCA342A, RRID: AB_2074557).

Techniques: Activation Assay, Isolation, Flow Cytometry, Expressing

Journal: OncoImmunology

Article Title: A targeted MAVS fusion protein for controlled innate immune activation and antitumor therapy

doi: 10.1080/2162402x.2025.2478850

Figure Lengend Snippet: Figure 3. TRIAS expression on tumor cells induced proinflammatory effects (a) the expression of TRIAS in MC38 cells was confirmed by flow cytometry. (b) Cell lysates were collected to detect the phosphorylation levels of IRF-3 and NF-κB following antigen stimulation at the indicated time points. (c) FCM analysis of MHC I and MHC II expression in MC38 cells engineered with different constructions. (d) The viability of the TRIAS cells exposed to different concentrations of antigens was detected by a CCK-8 assay.

Article Snippet: Detection of MHC molecules expression on tumor cells UTD, GFP or TRIAS MC38 cells were treated with 2 μg/mL EDB-his protein for 24 hours, the expression of MHC molecules was detected by FCM using APC-conjugated anti-mouse H-2Kb antibodies (Biolegend 116,517) and PerCP-conjugated anti-mouse MHC II (I-A/I-E) antibodies (Elabscience, E-ABF0990F).

Techniques: Expressing, Flow Cytometry, Phospho-proteomics, CCK-8 Assay