Journal: Cells

Article Title: MRGPRX2 Activation by Rocuronium: Insights from Studies with Human Skin Mast Cells and Missense Variants

doi: 10.3390/cells10010156

Figure Lengend Snippet: Rocuronium induces MRGPRX2 internalization in LAD2 cells and primary human skin MCs. ( A ) Flow cytometry histograms for MRGPRX2 cell surface expression (solid black line) and isotype (dotted gray line) are shown. Histograms are representative of three independent experiments. ( B ) The comparison of MRGPRX2 expression levels between human MC line LAD2 cells and primary human skin MCs. ( C ) LAD2 cells were stimulated with rocuronium (2 mg/mL) for 30 min, and MRGPRX2 internalization was determined by flow cytometry. ( D ) The adjusted MFI levels of MRGPRX2 cell surface expression are shown. ( E , F ) Rocuronium-induced MRGPRX2 internalization and adjusted MFI levels of MRGPRX2 cell surface expression were determined in primary human skin MCs. All data points are the mean ± SEM of at least three experiments. Statistical significance was determined by two-tailed unpaired t -test. * p < 0.05.

Article Snippet: Plasmid encoding hemagglutinin (HA)-tagged human MRGPRX2 in pReceiver-MO6 vector was obtained from GeneCopoeia (Rockville, MD, USA).

Techniques: Flow Cytometry, Expressing, Comparison, Two Tailed Test

Journal: Cells

Article Title: MRGPRX2 Activation by Rocuronium: Insights from Studies with Human Skin Mast Cells and Missense Variants

doi: 10.3390/cells10010156

Figure Lengend Snippet: MRGPRX2 mutations rendered the receptor unresponsiveness to rocuronium. ( A ) Snake diagram of MRGPRX2 indicating three missense mutations identified in the patient. ( B ) RBL-MRGPRX2 were stimulated with different concentrations of rocuronium for 30 min, and β-hexosaminidase release was determined. ( C ) Untransfected WT RBL-2H3 cells were stimulated with rocuronium (2 mg/mL) for 30 min, and β-hexosaminidase release was determined. ( D ) Cell surface expression of WT MRGPRX2 and its variants was determined by flow cytometry. ( E ) Cells expressing WT MRGPRX2 and its variants were exposed to buffer (control) or rocuronium (1 mg/mL) for 30 min, and β-hexosaminidase release was determined. ( F ) Dose response of rocuronium-induced β-hexosaminidase release was determined in cells expressing WT MRGPRX2 and M196I. Data are the mean ± SEM of at least three experiments. For comparisons of two samples, two-tailed unpaired t -test was used. For comparisons of multiple samples to a control group, one-way ANOVA with Dunnett’s post-hoc test was used. ** p < 0.01, *** p < 0.001 and **** p < 0.0001.

Article Snippet: Plasmid encoding hemagglutinin (HA)-tagged human MRGPRX2 in pReceiver-MO6 vector was obtained from GeneCopoeia (Rockville, MD, USA).

Techniques: Expressing, Flow Cytometry, Control, Two Tailed Test

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: ?-defensins activate human mast cells via Mas-related Gene-X2 (MrgX2)

doi: 10.4049/jimmunol.1300023

Figure Lengend Snippet: LAD2 mast cells were stably transduced with scrambled shRNA control lentivirus or shRNA lentivirus targeted against MrgX2. (A) Western blotting was performed to determine MrgX2 expression in control and MrgX2 knockdown (KD) cells. (B) shRNA control and MrgX2 KD cells were stimulated with hBD2, hBD3, cortistatin (CST) or C3a and percent degranulation (β-hexosaminidase release) was determined. Data are mean ± SEM of three experiments. Statistical significance was determined by one-way ANOVA with Bonferroni's post test. * indicates p<0.01 and ** indicates p<0.001.

Article Snippet: MrgX2 antibody was purchased from Novus Biologicals (Littleton, CO).

Techniques: Stable Transfection, Transduction, shRNA, Control, Western Blot, Expressing, Knockdown

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: ?-defensins activate human mast cells via Mas-related Gene-X2 (MrgX2)

doi: 10.4049/jimmunol.1300023

Figure Lengend Snippet: (A) RBL-2H3 cells stably expressing MrgX2 were stimulated with buffer, hBD2, hBD3 or cortistatin (CST) for 30 min and β-hexosaminidase release was measured. Data shown are representative of 3 similar experiments. Statistical significance was determined by one-way ANOVA with Bonferroni's post test. * indicates p<0.01 and ** indicates p<0.001. RBL-2H3 cells stably expressing MrgX2 were loaded with Indo-1AM and Ca2+ mobilization in response to (B) hBD2, (C) hBD3 or (D) CST was determined. HEK293 cells stably expressing MrgX2 were loaded with Indo-1AM and Ca2+ mobilization in response to (E) hBD3 or (F) CST was determined. Traces shown are representative of 3 individual experiments.

Article Snippet: MrgX2 antibody was purchased from Novus Biologicals (Littleton, CO).

Techniques: Stable Transfection, Expressing

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: ?-defensins activate human mast cells via Mas-related Gene-X2 (MrgX2)

doi: 10.4049/jimmunol.1300023

Figure Lengend Snippet: (A) BMMCs were transiently transfected with HA tagged MrgX2 (solid line) or control plasmid vector (broken line) and MrgX2 receptor expression level was analyzed using flow cytometry. A representative histogram is shown. (B) Control and MrgX2 expressing BMMCs were incubated with DNP specific mouse IgE (1 μg/mL, 16 h). Cells were exposed to buffer (control), CST, hBD3, mCRAMP or DNP-BSA (10 ng/mL) for 30 minutes and β-hexosaminidase release was measured. LAD2 cells were stimulated with mCRAMP and (C) intracellular Ca2+ mobilization or (D) degranulation was determined. Traces are representative of 3 independent experiments. Bar graphs represent mean ± SEM of three experiments. Statistical significance was determined by one-way ANOVA with Bonferroni's post test. * indicates p<0.01.

Article Snippet: MrgX2 antibody was purchased from Novus Biologicals (Littleton, CO).

Techniques: Transfection, Control, Plasmid Preparation, Expressing, Flow Cytometry, Incubation

Journal: Pharmacology Research & Perspectives

Article Title: The effect of substance P and its common in vivo‐formed metabolites on MRGPRX2 and human mast cell activation

doi: 10.1002/prp2.990

Figure Lengend Snippet: The C‐terminally truncated SP metabolite SP(1–9)‐COOH has ablated NK1R activation but retains MRGPRX2 activation. HEK293‐NK1R cells and HEK293‐MRGPRX2 cells were loaded with FURA‐2 and stimulated with a range of concentrations of peptides SP, SP(1–9)‐COOH, and SP(1–7)‐COOH. Peak calcium response ratios were plotted, or time courses generated. The parent peptide SP exhibited concentration‐dependent calcium mobilization response at both NK1R (A, B) and MRGPRX2 (C, D). The endogenous metabolites SP(1–9)‐COOH and SP(1–7)‐COOH showed no response at NK1R while only the former retained activity at MRGPRX2 (C, E, F). Responses were MRGPRX2‐dependent as mock‐transfected cells showed no response to SP and its metabolites (D, E, F). Data shown are means ± SEM, n = 7–9. Dotted lines represent the time of administration of peptides at t = 30 s.

Article Snippet: MRGPRX2 expressing HEK‐293 cells (HEK‐MRGPRX2) were generated through transient co‐transfection with a human MRGPRX2 construct (Genscript) and human Gα15 (cDNA Resource Center) to enhance GPCR coupling for detection of Ca 2+ mobilization.

Techniques: Activation Assay, Generated, Concentration Assay, Activity Assay, Transfection

Journal: Pharmacology Research & Perspectives

Article Title: The effect of substance P and its common in vivo‐formed metabolites on MRGPRX2 and human mast cell activation

doi: 10.1002/prp2.990

Figure Lengend Snippet: Both SP(1–9)‐COOH triggered calcium mobilization and MC degranulation are significantly ablated in MRGPRX2‐KD LAD2 cells. SP and SP(1–9)‐COOH activation of both Ca 2+ mobilization (A) and degranulation (B) were strongly inhibited by MRGPRX2 knockdown. (A) LAD2 cells were loaded with FURA‐2 and then stimulated with peptides and peak calcium ratios measured. Data shown are means ± S.E.M., n = 6–7. (B) Mast cell degranulation was measured by quantifying β‐hexosaminidase release, which is shown as a % of total cellular release. Data shown are means ± S.E.M., n = 6–7. Activation of the IgE‐dependent pathway (NIP‐BSA [30 ng/ml]) was unaffected by MRGPRX2 knockdown. For statistical analysis, a two‐way ANOVA was conducted with Bonferroni multiple comparison post hoc test. Each MRGPRX2 KD response was compared to the respective response in the LAD2 WT cells. ** p < .001, *** p < .0005, **** p < .000005.

Article Snippet: MRGPRX2 expressing HEK‐293 cells (HEK‐MRGPRX2) were generated through transient co‐transfection with a human MRGPRX2 construct (Genscript) and human Gα15 (cDNA Resource Center) to enhance GPCR coupling for detection of Ca 2+ mobilization.

Techniques: Activation Assay, Knockdown, Comparison

Journal: Pharmacology Research & Perspectives

Article Title: The effect of substance P and its common in vivo‐formed metabolites on MRGPRX2 and human mast cell activation

doi: 10.1002/prp2.990

Figure Lengend Snippet: C‐terminal amidation of SP(1–9) and SP(1–7) enhances their ability to activate MRGPRX2. (A) LAD2 cells were loaded with FURA‐2 and then stimulated with SP peptides. Peak calcium responses were measured. Data shown are means ± S.E.M., n = 6–8. (B) LAD2 cells were stimulated with SP peptides and degranulation quantified. Data shown are means ± S.E.M., n = 5–6.

Article Snippet: MRGPRX2 expressing HEK‐293 cells (HEK‐MRGPRX2) were generated through transient co‐transfection with a human MRGPRX2 construct (Genscript) and human Gα15 (cDNA Resource Center) to enhance GPCR coupling for detection of Ca 2+ mobilization.

Techniques:

Journal: Cells

Article Title: Small-Molecule Host-Defense Peptide Mimetic Antibacterial and Antifungal Agents Activate Human and Mouse Mast Cells via Mas-Related GPCRs

doi: 10.3390/cells8040311

Figure Lengend Snippet: smHDPMs activate human mast cells via MRGPRX2. ( A ) LAD2 cells pretreated with or without pertussis toxin (PTx, 100 ng/mL, 16h) were exposed to vehicle (Control) or smHDPMs compounds (Cpd) 1, 2, and 3 (3 µM each) for 30 min and percentage of β-hexosaminidase release was measured. ( B ) Western blotting was performed to determine the expression level of MRGPRX2 in Control and MRGPRX2 knockdown LAD2 cells. ( C ) Control and knockdown cells were stimulated with smHDPMs Cpd 1, 2, and 3 (3 µM each) and percentage of β-hexosaminidase release was determined. All the points are expressed as a mean ± SEM of three experiments in triplicate. Statistical significance was determined by two-tailed unpaired t-Test. *** indicates p value <0.001,** indicates p value <0.01 and * indicates p value <0.05.

Article Snippet: Polyclonal MRGPRX2 Ab was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Control, Western Blot, Expressing, Knockdown, Two Tailed Test

Journal: Cells

Article Title: Small-Molecule Host-Defense Peptide Mimetic Antibacterial and Antifungal Agents Activate Human and Mouse Mast Cells via Mas-Related GPCRs

doi: 10.3390/cells8040311

Figure Lengend Snippet: smHDPMs induce degranulation in RBL-2H3 cells expressing MRGPRX2. ( A ) Untransfected RBL-2H3 cells (RBL-2H3) and cells stably expressing MRGPRX2 (RBL-2H3-MRGPRX2) were exposed to vehicle (Control) or smHDPMs, (Cpd 1, 2, and 3, 3 µM) for 30 min and percentage of β-hexosaminidase release was determined. Concentration–response curves for ( B ) Cpd 1, ( C ) Cpd 2, and ( D ) Cpd 3 were determined using RBL-2H3-MRGPRX2 cells. Data are presented as a mean ± SEM and are representative of three independent experiments in triplicate. Statistical significance was determined by two-tailed unpaired t-Test. *** indicates p value <0.001.

Article Snippet: Polyclonal MRGPRX2 Ab was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing, Stable Transfection, Control, Concentration Assay, Two Tailed Test

Journal: Cells

Article Title: Small-Molecule Host-Defense Peptide Mimetic Antibacterial and Antifungal Agents Activate Human and Mouse Mast Cells via Mas-Related GPCRs

doi: 10.3390/cells8040311

Figure Lengend Snippet: Compound 4 and Compound 5 induce degranulation in RBL-2H3 cells expressing MRGPRX2. ( A ) Untransfected RBL-2H3 cells (RBL-2H3) and cells stably expressing MRGPRX2 (RBL-2H3-MRGPRX2) were exposed to vehicle (Control), smHDPMs compound (Cpd) 4, and Cpd 5 (3 µM each) for 30 min and percentage of β-hexosaminidase release was determined. Concentration–response curves for ( B ) Cpd 4 and ( C ) Cpd 5 were performed using RBL-2H3-MRGPRX2 cells. Data are presented as mean ± SEM and are representative of three independent experiments in triplicate. Statistical significance was determined by two-tailed unpaired t-Test. *** indicates p value <0.001.

Article Snippet: Polyclonal MRGPRX2 Ab was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Expressing, Stable Transfection, Control, Concentration Assay, Two Tailed Test

Journal: Cells

Article Title: Small-Molecule Host-Defense Peptide Mimetic Antibacterial and Antifungal Agents Activate Human and Mouse Mast Cells via Mas-Related GPCRs

doi: 10.3390/cells8040311

Figure Lengend Snippet: Missense MRGPRX2 variants do not respond to smHDPMs for degranulation. ( A ) RBL-2H3 cells were transiently transfected with cDNA encoding MRGPRX2 or its missense variants D184H and G165E. Receptor expression was determined by flow cytometry using anti-MRGPRX2-PE antibody. ( B ) RBL-2H3 expressing MRGPRX2 variants (D184H and G165E) were stimulated with smHDPMs (Cpds 1–5, 3.0 µM) for 30 min and β-hexosaminidase release was determined. All the points are expressed as a mean ± SEM of three experiments in triplicate. Statistical significance was determined by one-way ANOVA. **** indicates p value <0.0001,*** indicates p value <0.001 and ** indicates p value <0.01.

Article Snippet: Polyclonal MRGPRX2 Ab was purchased from Novus Biologicals (Littleton, CO, USA).

Techniques: Transfection, Expressing, Flow Cytometry

Journal: iScience

Article Title: The MRGPRX2-substance P pathway regulates mast cell migration

doi: 10.1016/j.isci.2024.110984

Figure Lengend Snippet: MRGPRX2 expression is highly heterogeneous between donors during human mast cell differentiation and heterogeneity is maintained in mature mast cells (A) Expression of MC markers and MRGPRX2 during in vitro differentiation of peripheral blood derived human MCs. Flow cytometric analysis using fluorescently labeled antibodies to quantify marker expression. Data are specific geometric mean fluorescence (sGMFI) (specific antibody – control values) or % of positive cells (percentage of cells with fluorescent values above control) for n = 3–12 separate donors (as shown) at day 0, 1, 3, 5, 7, and 9 weeks. (B) MRGPRX2 heterogeneity in comparison to Siglec6. Histograms showing Siglec6 or MRGPRX2 fluorescence intensity (x axis) vs. cell count (y axis) indicative of marker expression in three separate donors at maturity. Siglec6 fluorescence is shown in the top panels with homogenous populations of immuno-positive cells (purple) vs. control (turquoise). MRGPRX2 expression is shown in the lower panels with heterogeneous numbers of cells showing MRGPRX2-specific fluorescence intensity (purple) greater than control (turquoise).

Article Snippet: Sections were subsequently incubated with primary antibodies specific to tryptase (clone: EPR9522, 1:4000, Abcam, Cambridge, UK), and MRGPRX2 (clone 1D3, 1:100, Origene, Rockville, USA) diluted in blocking buffer for an hour at room temperature.

Techniques: Expressing, Cell Differentiation, In Vitro, Derivative Assay, Labeling, Marker, Fluorescence, Control, Comparison, Cell Counting

Journal: iScience

Article Title: The MRGPRX2-substance P pathway regulates mast cell migration

doi: 10.1016/j.isci.2024.110984

Figure Lengend Snippet: Substance P selectively activates MRGPRX2 + mast cells inducing rapid degranulation which is inhibited by compound A (A) Representative dot plots showing degranulation in mature MCs, as measured by CD63 externalization (x axis), vs. MRGPRX2 expression (y axis) after 15 min of treatment with Substance P (SP) (1 μM), or IgE/αIgE (100 ng/1 μg mL −1 ). Percentage of immuno-positive cells in each quadrant is shown for stimulated cells in black text. Unstimulated control percentages are shown in gray italics. (B) Time course of SP or IgE/αIgE induced degranulation (as indicated, 0–30 min), measured by flow cytometric analysis of CD63 externalization bound by CD63-specific fluorescent antibody. Data are mean ± SD of n = 3 from selected high MRGPRX2 expressing donors. Significant differences vs. time 0 are indicated (∗/† = p < 0.05, ∗∗/† † = p < 0.01), two-way ANOVA with Sidak’s multiple comparison post-test. (C) Substance P-induced degranulation as measured by CD63 externalization in conjunction with beta-hexosaminidase release (0.03–30 μM SP; left panel) and membrane expression of MRGPRX2 (0.1–100 μM SP; right panel) Data are mean ± SD of n = 3 replicates from mix of 3 donors p < 0.0001 two-way ANOVA with Dunnet’s multiple comparison post-test (left panel) or mean ± SEM of n = 3 from 3 donors (EC50 calculated by non-linear regression, three parameter best-fit curve; right panel). (D) Representative dot plots from three donors showing degranulation, as measured by flow cytometric analysis of immuno-specific CD63 externalization (x axis), vs. MRGPRX2 expression (y axis) after pretreatment with 10 μM compound A or vehicle control, for 30 min followed by 15 min of treatment with SP (3 μM), or vehicle control. Percentage of cells in each quadrant is shown for stimulated cells. Fluorescence Minus One (FMO) controls, with absence of specific antibody, for gating are shown in Figure S1 . (E) Degranulation of MCs as measured by flow cytometric analysis of immuno-specific CD63 externalization (top panel) or beta-hexosaminidase enzyme release (middle panel) after pretreatment with 100 μM Compound A or vehicle control for 30 min, followed by stimulation with SP (10 μM), C3a (50 nM), or vehicle control for 30 min. Comparison between SP and C3a ligand induced degranulation on a per-donor basis is shown (lower panel). Data are n = 7 from 7 separate donors. p values as indicated (two-way ANOVA with Sidak’s multiple comparison post-test). (F) Inhibition of degranulation induced by 0.3, 1, or 3 μM SP after pretreatment with compound A (3 nM–100 μM). Data are mean ± SEM of n = 5 separate donors. Lines represent variable slope least squares fit, with calculated IC 50 values labeled.

Article Snippet: Sections were subsequently incubated with primary antibodies specific to tryptase (clone: EPR9522, 1:4000, Abcam, Cambridge, UK), and MRGPRX2 (clone 1D3, 1:100, Origene, Rockville, USA) diluted in blocking buffer for an hour at room temperature.

Techniques: Expressing, Control, Comparison, Membrane, Fluorescence, Inhibition, Labeling

Journal: iScience

Article Title: The MRGPRX2-substance P pathway regulates mast cell migration

doi: 10.1016/j.isci.2024.110984

Figure Lengend Snippet: Substance P induced chemotaxis of immature and mature MCs and aids long term selection of MRGPRX2 + MC from progenitors (A) Representative dot plots showing migrated CD117+ immature MC recovered from the lower wells of the chemotaxis chamber after chemotaxis to 0–10 μM SP chemotaxis gradient, or equimolar (10 μM) SP on both sides (chemoattractant control). Cells were recovered and analyzed by counting the number of CD117-specific fluorescently labeled cells using flow cytometry (y axis) within the size parameter for MCp (SSC-A). The number of CD117+ cells is shown on the lower right of each plot. Plots show SSC-A (side scatter area) on the y axis and CD117 on the x axis. (B) Example of donor specific chemotaxis to SP in immature MC (donor 1–3) or mature MCs (donor 4–6). Number of migrating cells immuno-positive for CD117 (y axis) to each concentration of SP (x axis) as counted by flow cytometry is shown. (C) Inhibition of chemotaxis in immature (left) and mature (right) MCs in response to SP (0.05–10 μM [x axis]) in the presence of vehicle control (purple) or compound A (mauve). Upper panels show examples of single donor migration to SP. Lower panels are normalized mean data expressed as a percentage of the maximum number of migrated cells and are mean ± SEM of 6 (left) or 4 (right) experiments from separate donors. ∗∗ = p < 0.01 (two-way ANOVA with Sidak’s multiple comparison post-test). (D–N) Exposure of MCp to SP in cell culture medium during week 0–4 of differentiation. MCp were exposed to 10 μM SP added to media as cells were routinely cultured and media replenished. MC markers as labeled (D–K) and cell viability (L), total cell count (M), and cell density (N) are shown. Data are mean ± SEM of 3 donors. ∗ = p < 0.05, ∗∗ = p < 0.01 (two-way ANOVA with Sidak’s multiple comparison post-test).

Article Snippet: Sections were subsequently incubated with primary antibodies specific to tryptase (clone: EPR9522, 1:4000, Abcam, Cambridge, UK), and MRGPRX2 (clone 1D3, 1:100, Origene, Rockville, USA) diluted in blocking buffer for an hour at room temperature.

Techniques: Chemotaxis Assay, Selection, Control, Labeling, Flow Cytometry, Concentration Assay, Inhibition, Migration, Comparison, Cell Culture, Cell Counting

Journal: iScience

Article Title: The MRGPRX2-substance P pathway regulates mast cell migration

doi: 10.1016/j.isci.2024.110984

Figure Lengend Snippet: MRGPRX2 + mast cells are increased in inflammatory skin conditions and locate in proximity of SP + nerve fibers (NFs) during skin reinnervation in vitro (A) Representative images of healthy and psoriasis skin sections taken from skin punch biopsies, stained with anti-sera to MRGPRX2 (cyan) and MC tryptase (magenta) with DAPI (blue) nuclear counterstain. Areas of colocalization of MRGPRX2 and tryptase are indicated by arrows. MRGPRX2 negative MC are marked by +. Scale bar: 20 μm. (B and C) Density (B) and percentage (C) of MRGPRX2+ MCs in healthy and psoriasis skin sections. Cell numbers were obtained by manual counting of images as described in the section. Data are mean ± SEM of n = 6 donors. ns = not significant (unpaired t test). (D) Correlation between psoriasis area and severity index (PASI) and MRGPRX2+ MC density in psoriasis skin sections, measured by Pearson’s correlation coefficient. (E–G) Number of (N°) tryptase + MCs (E), tryptase + /MRGPRX2 + MCs (F), and percentage of (%) tryptase positive MCs (G) which are MRGPRX2 + during and after the course of 10 days of culture for skin reinnervation/nerve sprouting. Cell numbers were obtained by manual counting of images as described in the section. Of note: all the layers of the skin are reinnervated by day 5. Data are mean ± SEM of n = 4 sections (dermis and epidermis on each section were imaged in whole at 200× and stitched for analysis) from n = 2 biopsies from 1 donor. (H) Representative images of human skin biopsies after 2, 4, 7, and 10 days of co-culture with sensory neurons and stained with anti-sera to PGP9.5, tryptase and MRGPRX2 and counterstained with DAPI nuclear stain. Scale bar: 50 (high magnification insert) or 100 μm. (I–K) Number and percentage of tryptase + MCs in proximity to PGP9.5 + or SP + NFs (within 20 μm from NFs) during and after the course of skin culture reinnervation/nerve sprouting. Of note: all the layers of the skin are reinnervated by day 5. Data are mean ± SEM of n = 4 sections from n = 2 biopsies from 1 donor.

Article Snippet: Sections were subsequently incubated with primary antibodies specific to tryptase (clone: EPR9522, 1:4000, Abcam, Cambridge, UK), and MRGPRX2 (clone 1D3, 1:100, Origene, Rockville, USA) diluted in blocking buffer for an hour at room temperature.

Techniques: In Vitro, Staining, Co-Culture Assay

Journal: iScience

Article Title: The MRGPRX2-substance P pathway regulates mast cell migration

doi: 10.1016/j.isci.2024.110984

Figure Lengend Snippet:

Article Snippet: Sections were subsequently incubated with primary antibodies specific to tryptase (clone: EPR9522, 1:4000, Abcam, Cambridge, UK), and MRGPRX2 (clone 1D3, 1:100, Origene, Rockville, USA) diluted in blocking buffer for an hour at room temperature.

Techniques: Plasmid Preparation, Recombinant, Staining, Chemotaxis Assay, Migration, Software