Journal: The American Journal of Pathology

Article Title: Microglia Promote Neurodegeneration and Hyperkatifeia during Withdrawal and Abstinence from Binge Alcohol

doi: 10.1016/j.ajpath.2025.10.005

Figure Lengend Snippet: Gi designer receptors exclusively activated by designer drugs (DREADD) inhibition of microglia blocks alcohol-induced neuronal cell death. A – D: Organotypic brain slice cultures (OBSCs) from CX3CR1.Cre ERT2 .hM4di.mCitrine (+/–) mice were used to assess the role of microglia on neuronal death ex vivo. A: Representative image with or without addition of 4-hydroxy-tamoxifen (4-OH-TAM; 10 ng/mL) to induce Cre recombination, which caused a robust increase in mCitrine within ionized calcium-binding adapter molecule 1 (Iba-1) + microglia, consistent with microglial DREADD expression. Arrowheads denote colocalization between Iba-1 and mCitrine. B: Representative images of propidium iodide (PI) labeling of cell death in OBSC in response to ethanol. C: Ethanol (EtOH; 100 mmol/L; 4 days) caused a robust increase in cell death 24 hours into withdrawal, which was significantly blocked by microglial Gi signaling induced by clozapine N-oxide [CNO; two-way analysis of variance (ANOVA) main effect of ethanol F 1,20 = 11.37, P = 0.003; Sidak post test]. D and E: Induction of hM4Di Gi DREADD inhibition of microglia in vivo . CX3CR1.Cre ERT2 .hM4Di (+/–) mice received 10 days of water or binge ethanol (5 g/kg per day; intragastric) with or without CNO (3 mg/kg; intraperitoneally; 10 hours after ethanol). Assessments were performed 24 hours following the final treatment. D: Representative images of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) cell death stain in orbitofrontal cortex. E: Ethanol increased TUNEL + cell death by approximately threefold, which was prevented by Gi DREADD inhibition of proinflammatory microglia (two-way ANOVA, EtOH × CNO interaction F 1,16 = 8.5, P = 0.01; Sidak post test). N = 3 control and CNO slices ( C ); N = 10 ethanol slices ( C ); N = 8 ethanol + CNO slices ( C ); N = 4 water + phosphate-buffered saline (PBS) mice and water + CNO mice ( D and E ); N = 6 ethanol + PBS mice and ethanol + CNO mice ( D and E ). ∗ P < 0.05, ∗∗ P < 0.01. Scale bars: 20 μm ( A ); 500 μm ( B ); 25 μm ( D ).

Article Snippet: For Gi DREADD inhibition, CX3CR1.Cre ERT2 .hM4di mice were bred in-house from CX3CR1.Cre ERT2 (JAX strain 020940; 129P2 background) and DIO.hm4di mice (JAX strain 026219; 129S background) obtained from Jackson Laboratory.

Techniques: Inhibition, Slice Preparation, Ex Vivo, Binding Assay, Expressing, Labeling, In Vivo, End Labeling, TUNEL Assay, Staining, Control, Saline

Journal: The American Journal of Pathology

Article Title: Microglia Promote Neurodegeneration and Hyperkatifeia during Withdrawal and Abstinence from Binge Alcohol

doi: 10.1016/j.ajpath.2025.10.005

Figure Lengend Snippet: Gi designer receptors exclusively activated by designer drugs inhibition of microglia fails to attenuate alcohol-induced negative affect in males. A: Experimental design. Male CX3CR1.Cre ERT2 .hM4Di (+/–) mice received 10 days of water or binge ethanol (EtOH; 5 g/kg per day; intragastric) with or without clozapine N-oxide (CNO) 10 hours after treatment. Mice underwent open field testing for anxiety-like behavior 3 days into withdrawal and conditioned fear for acquisition and extinction at 34 days into abstinence. B: hM4di expression was measured by RT-PCR. Males showed approximately 25% of the level hM4di expression as females. t- Test was used. C and D: Microglial inhibition failed to prevent the induction of anxiety-like behavior caused by alcohol seen change in the distance traveled in the center of the open field [one-way analysis of variance (ANOVA) F 2,18 = 2.0, P = 0.16; C ] and a reduction in the time spent in the center of the open field (one-way ANOVA F 2,17 = 3.99, P = 0.038; Sidak post test; D ). E: Fine motor movement was significantly reduced in the EtOH + CNO group (one-way ANOVA F 2,18 = 3.5, P = 0.05; Sidak post test). F: Ethanol had no impact on conditioned fear memory acquisition. G: Neither ethanol nor microglial inhibition had any effect on contextual fear memory (two-way ANOVA time × treatment F 8,72 = 0.9, P = 0.5). H: Microglial inhibition had no impact on fear memory extinction (one-way repeated measures ANOVAs: control F 2.86,16.19 = 3.2, P = 0.053; ethanol F 2.999,17.99 = 0.38, P = 0.77; ethanol + CNO F 1.94,11.7 = 0.9, P = 0.43). I: No robust proinflammatory gene induction signature was seen with ethanol in the hM4di mice, with little impact of microglial inhibition. ∗ P < 0.05. GFAP, glial fibrillary acidic protein; NS, not significant; TLR, toll-like receptor; TNF, tumor necrosis factor.

Article Snippet: For Gi DREADD inhibition, CX3CR1.Cre ERT2 .hM4di mice were bred in-house from CX3CR1.Cre ERT2 (JAX strain 020940; 129P2 background) and DIO.hm4di mice (JAX strain 026219; 129S background) obtained from Jackson Laboratory.

Techniques: Inhibition, Expressing, Reverse Transcription Polymerase Chain Reaction, Control

Journal: The American Journal of Pathology

Article Title: Microglia Promote Neurodegeneration and Hyperkatifeia during Withdrawal and Abstinence from Binge Alcohol

doi: 10.1016/j.ajpath.2025.10.005

Figure Lengend Snippet: Gi designer receptors exclusively activated by designer drugs (DREADD) inhibition of microglia attenuates alcohol-induced negative affect in females. A: Experimental design. Female CX3CR1.Cre ERT2 .hM4Di (+/–) mice received 10 days of water or binge ethanol (EtOH; 5 g/kg per day; intragastric) with or without clozapine N-oxide (CNO) 10 hours after treatment. Mice underwent open field testing for anxiety-like behavior 3 days into withdrawal and conditioned fear for acquisition and extinction at 34 days into abstinence. B and C: Microglial inhibition prevented the induction of anxiety-like behavior caused by alcohol seen as a reduction in the distance traveled in the center of the open field [one-way analysis of variance (ANOVA) F 2,20 = 3.95, P = 0.036; Sidak post test; B ] and reduced time spent in the center of the open field (one-way ANOVA F 2,20 = 7.1, P = 0.0046; Sidak post test; C ). D: Microglial inhibition prevented the trend toward a reduction in fine motor movement in the open field caused by ethanol (one-way ANOVA F 2,21 = 2.16, P = 0.14). E: Ethanol had no impact on conditioned fear learning acquisition. F: In contextual fear memory assessments, ethanol increased freezing in the last minute of assessment that was prevented by microglial inhibition with CNO (two-way ANOVA time × treatment F 8,84 = 3.7, P < 0.0001; Sidak post test). G: Ethanol disrupted the extinction of conditioned fear memory, which was restored by microglial inhibition (one-way repeated measures ANOVAs: control F 2.4,13.5 = 2.1, P = 0.16; ethanol F 2.8,19.8 = 0.9, P = 0.45; ethanol + CNO F 3.0,17.6 = 6.6, P = 0.0035). H: The persistent induction of multiple proinflammatory genes in the cortex 5 weeks into abstinence was prevented by Gi DREADD inhibition of microglia (two-way ANOVA F 2,231 = 6.921, P = 0.001). I: A negative correlation between IFNA gene expression and time spent in the center of the open field was found. IFN gene expression failed a normality test; thus, a Spearman correlation coefficient was calculated. R = -0.45, P = 0.039. N = 8 female water + vehicle, ethanol + vehicle, and ethanol + CNO ( B – I ). ∗ P < 0.05, ∗∗ P < 0.01. GFAP, glial fibrillary acidic protein; NS, not significant; TNF, tumor necrosis factor.

Article Snippet: For Gi DREADD inhibition, CX3CR1.Cre ERT2 .hM4di mice were bred in-house from CX3CR1.Cre ERT2 (JAX strain 020940; 129P2 background) and DIO.hm4di mice (JAX strain 026219; 129S background) obtained from Jackson Laboratory.

Techniques: Inhibition, Control, Gene Expression

Journal: Cell death and differentiation

Article Title: Inhibition of inflammatory osteoclasts accelerates callus remodeling in osteoporotic fractures by enhancing CGRP + TrkA + signaling.

doi: 10.1038/s41418-024-01368-5

Figure Lengend Snippet: Fig. 1 Upregulation of Cx3cr1+iOCs in the bone callus is associated with diminished trabecular formation during the remodeling of osteoporotic fractures. A Schematic of the experimental design to observe the role of Cx3cr1+iOCs in callus remodeling during osteoporotic fractures. B Representative Micro-CT images of femoral fractures at 2- and 4-weeks post-fracture. Scale bar: 1 mm. Representative HE (C) and SOFG (D) stained images of femoral fractures in Sham and OVX mice at 2- and 4-weeks post-fracture. Upper panels show global views; lower panels provide close-up views of the fracture sites. Tb trabecular bone. Scale bar: 1 mm. E Representative Cx3cr1 and Trap coimmunostaining images of calluses from Sham and OVX mice at 2- and 4-weeks post-fracture. ca callus. White dashed lines outline the callus boundaries, and white arrows point to Cx3cr1+Trap+ cells (Cx3cr1+iOCs). Scale bar: 200 μm. F–H Micro-CT quantitative analysis of the callus from (B). Parameters include bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N). n = 6. I Quantification of Cx3cr1+Trap+-positive osteoclasts (Cx3cr1+iOCs) in the callus from (E). n = 3. Data presented as mean ± SEM. **P < 0.01, ***P < 0.001 and ****P < 0.0001.

Article Snippet: Following this, the cells are stained with specific fluorescent dyes using antibodies targeting Cx3cr1 (1:100, NBP2-68616, Novus) and Ly6C (1:100, 88920, Cell Signaling Technology).

Techniques: Micro-CT, Staining

Journal: Cell death and differentiation

Article Title: Inhibition of inflammatory osteoclasts accelerates callus remodeling in osteoporotic fractures by enhancing CGRP + TrkA + signaling.

doi: 10.1038/s41418-024-01368-5

Figure Lengend Snippet: Fig. 2 Conditional knockout of Cx3cr1+iOCs accelerates callus remodeling in OVX mice. A Schematic of the experimental design to assess the effects of Cx3cr1+iOCs deficiency on callus remodeling in osteoporotic fractures. B Representative Micro-CT images of femoral fractures in Cx3cr1-Cre; DTRfl/flmice and WT littermates at 4 weeks post-fracture. Scale bar: 1 mm. Representative HE (C) and SOFG (D) stained images of femoral fractures in Cx3cr1-Cre; DTRfl/flmice and WT littermates at 4 weeks post-fracture. Upper panels display global views; lower panels provide close-ups of the fracture sites. Tb trabecular bone. Scale bar: 1 mm. E Immunofluorescence staining images showing osteocalcin (OCN) in calluses of Cx3cr1-Cre; DTRfl/flmice and WT littermates at 4 weeks post-fracture. ca callus. White dashed lines outline the callus boundaries. Scale bar: 200 μm. F–H Micro-CT quantitative analysis of the callus from (B). Parameters include bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N). n = 6. I Quantitative analysis of OCN fluorescence intensity in callus from (E). n = 3. Data presented as mean ± SEM. *P < 0.05, **P < 0.01 and ***P < 0.001.

Article Snippet: Following this, the cells are stained with specific fluorescent dyes using antibodies targeting Cx3cr1 (1:100, NBP2-68616, Novus) and Ly6C (1:100, 88920, Cell Signaling Technology).

Techniques: Knock-Out, Micro-CT, Staining

Journal: Cell death and differentiation

Article Title: Inhibition of inflammatory osteoclasts accelerates callus remodeling in osteoporotic fractures by enhancing CGRP + TrkA + signaling.

doi: 10.1038/s41418-024-01368-5

Figure Lengend Snippet: Fig. 3 In osteoporotic fractures, attenuation of CGRP+TrkA+ signaling correlates with upregulated Cx3cr1+iOCs expression in the callus, and the conditional knockout of Cx3cr1+iOCs enhances CGRP+TrkA+ signaling. A, B Immunofluorescence staining images and quantification of TUBB3 protein levels in calluses from Cx3cr1-Cre; DTRfl/flmice and WT littermates at 4 weeks post-fracture. C, D Immunofluorescence staining images and quantification of CGRP protein levels in calluses from Cx3cr1-Cre; DTRfl/flmice and WT littermates at 4 weeks post-fracture. E, F Immunofluorescence staining images and quantification of TrkA protein levels in calluses from Cx3cr1-Cre; DTRfl/flmice and WT littermates at 4 weeks post-fracture. White dashed lines outline the callus boundaries. ca callus. Scale bar: 200 μm. n = 3/group; data presented as mean ± SEM. **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Following this, the cells are stained with specific fluorescent dyes using antibodies targeting Cx3cr1 (1:100, NBP2-68616, Novus) and Ly6C (1:100, 88920, Cell Signaling Technology).

Techniques: Expressing, Knock-Out, Staining

Journal: Cell death and differentiation

Article Title: Inhibition of inflammatory osteoclasts accelerates callus remodeling in osteoporotic fractures by enhancing CGRP + TrkA + signaling.

doi: 10.1038/s41418-024-01368-5

Figure Lengend Snippet: Fig. 4 Sema3A secreted by Cx3cr1+iOCs inhibits CGRP+TrkA+ sensory nerve signaling via GSK3β–Akt pathway in vitro. A, B Flow cytometry analysis of Cx3cr1+Ly6C+ cells in bone marrow from sham and OVX mice. n = 3. C, D Representative coimmunostaining images of Cx3cr1 and Sema3A and quantification in calluses from Sham and OVX mice at 4 weeks post-fracture. ca callus. White arrows point to Cx3cr1+ Sema3A+ cells. Scale bar: 200 μm. n = 3. E Schematic of DRGs co-cultured under under Cx3cr1+iOCs conditions. F, G Immunofluorescence staining and quantification of TUBB3 in Sham, OVX, shRNA control, and shRNA Sema3A groups. Scale bar: 50 μm. n = 3. H–K Representative western blot images and quantification of CGRP, TrkA, p-TrkA, GSK3β, and p-GSK3β in DRGs treated with or without Sema3A from by cx3cr1+iOCs. n = 3. Data presented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Following this, the cells are stained with specific fluorescent dyes using antibodies targeting Cx3cr1 (1:100, NBP2-68616, Novus) and Ly6C (1:100, 88920, Cell Signaling Technology).

Techniques: In Vitro, Flow Cytometry, Cell Culture, Staining, shRNA, Control, Western Blot

Journal: Cell death and differentiation

Article Title: Inhibition of inflammatory osteoclasts accelerates callus remodeling in osteoporotic fractures by enhancing CGRP + TrkA + signaling.

doi: 10.1038/s41418-024-01368-5

Figure Lengend Snippet: Fig. 5 Following the conditional knockout of Sema3A in Cx3cr1+iOCs, callus remodeling in OVX mice accelerated, accompanied by enhanced CGRP+TrkA+ signaling. A Schematic of the experimental design to assess the effects of conditional deletion of Sema3A in Cx3cr1+iOCs on callus remodeling during osteoporotic fractures. B, C Representative HE (B) and Masson (C) stained images of femoral fractures in Cx3cr1-Cre; Sema3Afl/flOVX mice and their littermate controls at 4 weeks post-fracture. Upper panels display global views; lower panels show close-up views of the fracture sites. Tb trabecular bone. Scale bar: 1 mm. D, F Immunofluorescence staining images and quantification of CGRP protein levels in calluses from Cx3cr1-Cre; Sema3Afl/flOVX mice and their littermate controls at 4 weeks post-fracture. ca callus. White dashed lines outline the callus boundaries. Scale bar: 200 μm. n = 3. E, G Immunofluorescence staining images and quantification of TrkA protein levels in calluses from Cx3cr1-Cre; Sema3Afl/flOVX mice and their littermate controls at 4 weeks post-fracture. “ca” denotes callus. White dashed lines outline the callus boundaries. Scale bar: 200 μm. n = 3. Data presented as mean ± SEM. *P < 0.05, ***P < 0.001.

Article Snippet: Following this, the cells are stained with specific fluorescent dyes using antibodies targeting Cx3cr1 (1:100, NBP2-68616, Novus) and Ly6C (1:100, 88920, Cell Signaling Technology).

Techniques: Knock-Out, Staining

Journal: Cell death and differentiation

Article Title: Inhibition of inflammatory osteoclasts accelerates callus remodeling in osteoporotic fractures by enhancing CGRP + TrkA + signaling.

doi: 10.1038/s41418-024-01368-5

Figure Lengend Snippet: Fig. 7 Elevated Cx3cr1+iOCs and diminished CGRP+TrkA+ signaling in osteoporotic fracture tissues samples. A–G Pathological analysis of a femoral neck fracture in a 55-year-old male. Representative X-ray (A), H&E staining (B), and SO-FG staining (C) images are shown on the left. Immunofluorescence images of Cx3cr1 and Sema3A coimmunostaining (D), TUBB3 protein (E), CGRP protein (F), and TrkA protein (G) are shown on the right. Scale bar: 1 mm. H–N Pathological analysis of a femoral neck fracture in an 88-year-old female. Representative X-ray (H), H&E staining (I), and SOFG staining (J) images are shown on the left. Immunofluorescence images of Cx3cr1 and Sema3A coimmunostaining (K), TUBB3 protein (L), CGRP protein (M), and TrkA protein (N) are shown on the right. Scale bar: 1 mm. O–R Quantitative analysis of fluorescence intensity for Cx3cr1+Sema3A+, TUBB3, CGRP, and TrkA in fracture tissue samples from (D–G) and (K–N). n = 3/group; data presented as mean ± SEM. *P < 0.05, **P < 0.01 and ***P < 0.001.

Article Snippet: Following this, the cells are stained with specific fluorescent dyes using antibodies targeting Cx3cr1 (1:100, NBP2-68616, Novus) and Ly6C (1:100, 88920, Cell Signaling Technology).

Techniques: Staining

Journal: Journal of Ovarian Research

Article Title: Hypoxia-inducible factor-1α induces CX3CR1 expression and promotes the epithelial to mesenchymal transition (EMT) in ovarian cancer cells

doi: 10.1186/s13048-019-0517-1

Figure Lengend Snippet: CX3CR1 and HIF-1α expression by OvCa tissues: Ovarian tissues from normal and various cancer stages [well-differentiated (Stage I), moderately differentiated (Stage II), and poorly differentiated (Stage III)] were stained with anti-CX3CR1 and anti-HIF-1α antibodies. Magenta (AP) color shows CX3CR1, and Brown (DAB) color shows HIF-1α staining. An Aperio ScanScope CS system with a 40X objective captured digital images of each tissue. Representative cases are immuno-intensities of CX3CR1 and HIF-1α using image analysis Aperio ImageScope v.6.25 software

Article Snippet: After washing with PBS-T, tissue sections were incubated with streptavidin-horseradish peroxidase (HRP, Biolegend) (HIF-1α) or streptavidin-alkaline phosphatase (Jackson ImmunoResearch) (CX3CR1) and developed in diaminobenzidine and/or alkaline phosphatase red chromogen coloring agent.

Techniques: Expressing, Staining, Software

Journal: Journal of Ovarian Research

Article Title: Hypoxia-inducible factor-1α induces CX3CR1 expression and promotes the epithelial to mesenchymal transition (EMT) in ovarian cancer cells

doi: 10.1186/s13048-019-0517-1

Figure Lengend Snippet: Expression of CX3CR1 and CX3CL1 by OvCa cells. OVCAR-3, SW 626, and TOV-112D cells were stained with FITC-conjugated or anti-CX3CR1 and PE-conjugated isotype or anti-CX3CL1 antibodies, and the expressions were quantified by flow cytometry. The gray and white histograms represent isotypes and CX3CR1/CX3CL1 fluorescence intensity, respectively

Article Snippet: After washing with PBS-T, tissue sections were incubated with streptavidin-horseradish peroxidase (HRP, Biolegend) (HIF-1α) or streptavidin-alkaline phosphatase (Jackson ImmunoResearch) (CX3CR1) and developed in diaminobenzidine and/or alkaline phosphatase red chromogen coloring agent.

Techniques: Expressing, Staining, Flow Cytometry, Fluorescence

Journal: Journal of Ovarian Research

Article Title: Hypoxia-inducible factor-1α induces CX3CR1 expression and promotes the epithelial to mesenchymal transition (EMT) in ovarian cancer cells

doi: 10.1186/s13048-019-0517-1

Figure Lengend Snippet: Western blot expression of hypoxia regulatory markers in OvCa cells. Immunoblot detection of OVCAR-3, SW 626, and TOV-112D cells exposed to 3, 6, 9, or 12 h hypoxia respectively. Immunoblotting was accomplished with primary antibodies against HIF-1α, CX3CR1, EMT marker (Snail), and MMP-9. As an internal standard for equal loading, anti-GAPDH antibody was used to probe blots

Article Snippet: After washing with PBS-T, tissue sections were incubated with streptavidin-horseradish peroxidase (HRP, Biolegend) (HIF-1α) or streptavidin-alkaline phosphatase (Jackson ImmunoResearch) (CX3CR1) and developed in diaminobenzidine and/or alkaline phosphatase red chromogen coloring agent.

Techniques: Western Blot, Expressing, Marker

Journal: Journal of Ovarian Research

Article Title: Hypoxia-inducible factor-1α induces CX3CR1 expression and promotes the epithelial to mesenchymal transition (EMT) in ovarian cancer cells

doi: 10.1186/s13048-019-0517-1

Figure Lengend Snippet: mRNA analysis of hypoxia-induced signaling molecules in OvCa cells. Ovarian tumor cells (OVCAR-3, SW 626, and TOV-112D) were exposed to hypoxia for 3, 6, 9, or 12 h. Quantitative RT-PCR results for expressions of CX3CR1, HIF-1α, MMP9, Snail, Twist, N- and P-cadherin are shown. The data were normalized to the levels of the housekeeping gene (18 S), and the analyses were accomplished in triplicate. Data were presented as fold change in expression (± standard error); the asterisks indicate significant differences as determined by Student’s t -test (* P < 0.05; ** P < 0.01)

Article Snippet: After washing with PBS-T, tissue sections were incubated with streptavidin-horseradish peroxidase (HRP, Biolegend) (HIF-1α) or streptavidin-alkaline phosphatase (Jackson ImmunoResearch) (CX3CR1) and developed in diaminobenzidine and/or alkaline phosphatase red chromogen coloring agent.

Techniques: Quantitative RT-PCR, Expressing

Journal: Journal of Ovarian Research

Article Title: Hypoxia-inducible factor-1α induces CX3CR1 expression and promotes the epithelial to mesenchymal transition (EMT) in ovarian cancer cells

doi: 10.1186/s13048-019-0517-1

Figure Lengend Snippet: Migration and invasion induced by the CX3CR1/CX3CL1 interaction in hypoxic OvCa cells. OVCAR-3, SW 626, and TOV-112D cells were cultured in the presence of CoCl 2 (150 μM) (to mimic hypoxia), and with or without CX3CL1 (chemoattractant) and KC7F2 (inhibitor) in the medium. Over 6 days, migration and invasion were assessed. ( a ) Morphology and ( b ) quantitative analysis of surface area, representing the invading cells projecting out of the spheroid into the medium when using CX3CL1. For cells under hypoxia, there was minimal growth, or they remained as aggregates when treated with or without the inhibitor, KC7F2

Article Snippet: After washing with PBS-T, tissue sections were incubated with streptavidin-horseradish peroxidase (HRP, Biolegend) (HIF-1α) or streptavidin-alkaline phosphatase (Jackson ImmunoResearch) (CX3CR1) and developed in diaminobenzidine and/or alkaline phosphatase red chromogen coloring agent.

Techniques: Migration, Cell Culture

Journal: Cell Reports

Article Title: COX2 regulates senescence secretome composition and senescence surveillance through PGE 2

doi: 10.1016/j.celrep.2021.108860

Figure Lengend Snippet:

Article Snippet: Mouse anti-CX3CR1-164Dy , Fluidigm , # 3164023B, RRID: AB_2832247.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software, Sequencing