Journal:

Article Title: A novel and sensitive ELISA reveals that the soluble form of IL-13R-?2 is not expressed in plasma of healthy or asthmatic subjects

doi: 10.1111/j.1365-2222.2007.02921.x

Figure Lengend Snippet: sIL-13R-α2 detection in BAL fluid

Article Snippet: After static incubation for 2 h at 37 °C, plates were washed, and then incubated for 2 h, 37 °C, with biotinylated anti-human IL-13R-α2 (R&D Systems) diluted in PBS.

Techniques:

Journal: The Journal of Biological Chemistry

Article Title: Peptide-Major Histocompatibility Complex Dimensions Control Proximal Kinase-Phosphatase Balance during T Cell Activation

doi: 10.1074/jbc.M109.039966

Figure Lengend Snippet: T cell receptor triggering and effector responses are abrogated by pMHC elongation in a murine system. A , schematic representation of native and elongated versions of D b SCT containing the indicated membrane-proximal IgSF spacers, tethered to the plasma membrane by the native H-2D b stalk and transmembrane segments. B , surface expression levels of SCT constructs in TAP2-deficient CHO cells. C , primed transgenic T cells expressing the F5 TCR (10 4 T cells/well) were incubated with varying numbers of SCT-expressing CHO cells (denoted as APC/F5 ratio). Culture supernatants were assayed for IL-2 by ELISA after 24 h of incubation. A representative plot of three independent experiments is shown. The error bars represent ± S.D. D , F5/APC conjugates were incubated for 2 min at 37 °C before fixing and stained for phosphotyrosine and H-2D b (SCTs). The conjugates were imaged by confocal immunofluorescence microscopy (Fig. S1A ). Untransfected CHO cells were used to determine basal phosphorylation levels. Phosphotyrosine accumulation at the T cell interface was quantified as the ratio of interface/noninterface fluorescence ( black bars ). At least 25 conjugates were randomly chosen for analysis. Accumulation of SCTs was measured using a similar approach ( supplemental Fig. S1 B ). As a control, conjugates of B3Z hybridomas expressing the noncognate OT1 TCR and CHO cells expressing D b SCT were used. Accumulation of SCT at the interface was quantified for 19–25 randomly chosen conjugates for each SCT ( purple bars ). The error bars represent S.E. The dashed line represents enrichment ratio of 1 (no enrichment) E , fully differentiated F5 CTL were assessed for sensitivity to elongated pMHC expressed at high levels (as in B ) on CHO cells. Cytotoxic responses of F5 CTL to SCTs was tested in a 6-h 51 Cr release assay using CHO cells expressing native and maximally elongated (SCT-CD4) SCTs as targets. The data are representative of two independent experiments. The error bars represent ± S.D. F , cytotoxic responses by F5 CTL to low levels (∼50-fold lower than cells shown in B ) of SCT expression ( inset ). The data are representative of two independent experiments. G , cytolytic activity of F5 CTL and IL-2 release by F5 RAG −/− T cells in response to high levels of SCT expression (as in B ) in the presence of 0–10 μ m PP2 was measured as in C and E . The data are expressed as the percentages of reduction in response relative to responses in the absence of PP2. The concentration of PP2 that gave 50% inhibition (IC 50 ) was derived from error-weighted sigmoidal fitting of the data. The error bars represent ± S.D.

Article Snippet: For IL-2 measurements, culture supernatants and mouse recombinant IL-2 standards (Sigma) were incubated in microtitre plates coated with a capture anti-mouse IL-2 antibody (BD Pharmingen) for 2 h at 37 °C, washed, and incubated for another 1 h with a biotinylated detection anti-mouse IL-2 antibody (BD Pharmingen).

Techniques: Expressing, Construct, Transgenic Assay, Incubation, Enzyme-linked Immunosorbent Assay, Staining, Immunofluorescence, Microscopy, Fluorescence, Release Assay, Activity Assay, Concentration Assay, Inhibition, Derivative Assay

Journal: The Journal of Biological Chemistry

Article Title: Peptide-Major Histocompatibility Complex Dimensions Control Proximal Kinase-Phosphatase Balance during T Cell Activation

doi: 10.1074/jbc.M109.039966

Figure Lengend Snippet: Elongation of a human pMHC (gagSLY/HLA-A2) abrogates activation of G10 T cells expressing cognate TCR. A , schematic representation of single-chain constructs of native and elongated single-chain versions of HLA-A2/gag and corresponding coreceptor binding mutants ( yellow stars ). Constructs were stably expressed in TAP2-deficient CHO cells and sorted for comparable expression for use as surrogate antigen-presenting cells ( supplemental Fig. S3 A ). Also shown are soluble forms of native and elongated constructs comprising the extracellular portion followed by a C-terminal biotin acceptor site ( red dot ) and His 6 tag ( green line ). B , activation of primed G10 T cells was assessed by incubating 10 4 G10 cells/well with varying numbers of SCT-expressing APC (denoted as APC/G10 ratio) and IFNγ release measured by ELISA of culture supernatants after 8 h incubation. A representative plot of three independent experiments is shown. C , the human T cell clone SLY10 was used to assess responses to SCTs with mutated coreceptor-binding sites. Varying numbers of SCT-expressing CHO cells were incubated with 10 4 SLY10 cells/well, and MIP1b release was measured by ELISA of culture supernatants after 24 h of incubation. D , coreceptor binding to native and elongated SCTs was assessed by surface plasmon resonance (BIAcore). Monomeric biotinylated SCTs were immobilized by coupling to streptavidin conjugated CM5 flow cells (∼500 reference units). Binding curves were obtained by injecting CD8αα at a range of concentrations (0.9–154 μ m ) over flow cell surfaces immobilized with native or elongated SCTs. Immobilized biotinylated polyclonal anti-hamster IgG antibody was used as a control surface. The experiments were performed at 25 °C at a flow rate of 5 μl/min. Plateau response units were plotted against CD8αα concentration, and K D values were obtained by nonlinear curve fitting. The data are representative of two independent experiments. E , binding of native and elongated SCTs to G10 TCR was compared by surface plasmon resonance as described for D . The binding curves were obtained by injecting G10 TCR (0.1–7 μ m ) over immobilized SCTs, HLA-A2/NY-ESO, in addition to an irrelevant antibody-bound control surface.

Article Snippet: For IL-2 measurements, culture supernatants and mouse recombinant IL-2 standards (Sigma) were incubated in microtitre plates coated with a capture anti-mouse IL-2 antibody (BD Pharmingen) for 2 h at 37 °C, washed, and incubated for another 1 h with a biotinylated detection anti-mouse IL-2 antibody (BD Pharmingen).

Techniques: Activation Assay, Expressing, Construct, Binding Assay, Stable Transfection, Enzyme-linked Immunosorbent Assay, Incubation, SPR Assay, Concentration Assay

Journal: The Journal of Biological Chemistry

Article Title: Peptide-Major Histocompatibility Complex Dimensions Control Proximal Kinase-Phosphatase Balance during T Cell Activation

doi: 10.1074/jbc.M109.039966

Figure Lengend Snippet: TCR·CD3 triggering but not clustering is diminished by pMHC elongation. A , clean poly- l -lysine coated coverslips were incubated at 37 °C for 2 h with 50 μg/ml streptavidin/PBS. Biotinylated SCTs were immobilized on coverslips at comparable densities (Fig. S3 ). G10 cells were placed on SCT-coated coverslips and incubated for 1 min at 37 °C. The cells were fixed and permeabilized before serial staining CD3, Zap70, and CD45. The images were acquired using uniform settings between experiments. The cells adhered poorly on uncoated coverslips; therefore coverslips coated with biotinylated anti-HLA antibody was used as a control surface (without cognate TCR ligands). The scale bar represents 4 μm. B–D , the mean intensities for CD3, Zap70, and CD45 accumulation at the T cell interface was quantified. For a better appreciation of regions of TCR clustering pseudocolor scaled images of the CD3 distribution are shown (CD3 Scale) in A and E . The regions of CD3 clustering were arbitrarily defined by thresholding at two times the mean CD3 fluorescence intensity at the T cell interface (corresponding to the yellow regions in the pseudocolored scale images) and expressed as percentages of total CD3 interface fluorescence (Fig. S4 ). E , G10 cells were incubated with 10 μ m PP2 for 30 min prior to placing on to SCT-coated coverslips. Preparation and labeling were performed as described above. The scale bar represents 4 μm. The horizontal black bars represent the means. Statistical significance was determined by analysis of variance with correction for multiple comparisons. ns , p > 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: For IL-2 measurements, culture supernatants and mouse recombinant IL-2 standards (Sigma) were incubated in microtitre plates coated with a capture anti-mouse IL-2 antibody (BD Pharmingen) for 2 h at 37 °C, washed, and incubated for another 1 h with a biotinylated detection anti-mouse IL-2 antibody (BD Pharmingen).

Techniques: Incubation, Staining, Fluorescence, Labeling

Journal: Nature Communications

Article Title: Th1-poised naive CD4 T cell subpopulation reflects anti-tumor immunity and autoimmune disease

doi: 10.1038/s41467-025-57237-3

Figure Lengend Snippet: a Representative histogram of IL-7R, Adgre5, CD5 expression in IL-18R/Ly6C double-positive (DP) and double-negative (DN) naive CD4 T cells. b Representative dot plot of IL-18R and Ly6C expression in IL-7R sup-hi and IL-7R low naive CD4 T cells and memory-phenotype CD4 T cells (MP). c Representative dot plot of IL-18R/Ly6C DP population in naive CD4 T cells from each indicated tissue. d Tissue distribution of IL-18R/Ly6C DP population in naive CD4 T cells ( n = 10). e Representative histogram of naive CD4 T cells from spleen and thymus. f UMAP of murine naïve CD4 T cells from two different tissues (thymic CD4 SP and spleen). Cells were colored by cluster identity using label transfer of naïve CD4 T cell clusters from Fig. . g Stacked bar graph showing the proportion of naïve CD4 T cell clusters from thymus (Thy) and spleen (Spl). h Dot plots showing the distribution of pseudotime for the single cells in each cluster. i Alluvial plot to track the clonal population in thymus (Thy) and spleen (Spl) using TCR-seq data. j Box plots for the Shannon diversity index of TCR repertoire for each cluster in each tissue. k Expression of Ly6C and IL-18R in naive CD4 and CD8 single-positive (SP) cells in thymocytes stimulated with anti-CD3 for 3 days. l Proportion of Ly6C + cells in SP CD4 naive thymocytes stimulated by the indicated cytokines for 3 days ( n = 5). Statistical significance was confirmed by two-sided Mann-Whitney U -test with control group and each experimental group ( p = 0.0079). m Proportion of Ly6C + naive CD4 T cells in the splenocyte of WT, Stat1 -/- , Ifnar -/- , and Ifnar -/- Ifngr -/- mice (WT n = 5, Stat1 KO n = 5, Ifnar1 KO n = 3, Ifnar1 Ifngr1 KO n = 2). Data presented as the mean ± S.D. Statistical significance was confirmed by the two-sided Mann-Whitney U -test (* p < 0.05, ** p < 0.01, *** p < 0.001). Source data and exact statistics value are provided as a Source Data file.

Article Snippet: CD8 and NK depletion was performed by a single i.p injection of 200μg of anti-CD8 antibody (clone 2.43, BioXcell, #BE0061, lot 811521N1) and anti-NK1.1 antibody (clone PK136, BioXcell, #BE0036 lot 828622A2).

Techniques: Expressing, MANN-WHITNEY, Control

Journal: Scientific Reports

Article Title: IL-37 alleviates Coxsackievirus B3-induced viral myocarditis via inhibiting NLRP3 inflammasome-mediated pyroptosis

doi: 10.1038/s41598-022-22617-y

Figure Lengend Snippet: IL-37 alleviates myocarditis in CVB3-induced VMC mice. ( A , B ) M-mode echocardiographic images of mice in control group and VMC group and the LVEF, LVFS, IVSs, IVSd from the echocardiographic data. ( C ) The levels of cTnI in serum of mice in four groups were measured by ELISA. (Con: control group, * P < 0.01, ** P < 0.001, **** P < 0.00001, ns: not significant.)

Article Snippet: Anti-IL-1R8 (Santa Cruz, USA), anti-IL-37 (eBioscience, USA), and mouse IgG (Cell Signaling Technology, USA) were used.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: IL-37 alleviates Coxsackievirus B3-induced viral myocarditis via inhibiting NLRP3 inflammasome-mediated pyroptosis

doi: 10.1038/s41598-022-22617-y

Figure Lengend Snippet: IL-37 attenuated inflammatory cell infiltration and collagen tissue deposition. ( A ) Hearts of mice in all groups. ( B , D ) Histopathological changes in heart tissue of four groups were examined by H&E staining and Masson staining. ( C , E ) The pathologic score of mice in control group and VMC group. (*P < 0.01, **P < 0.001, ****P < 0.00001, ns: not significant.)

Article Snippet: Anti-IL-1R8 (Santa Cruz, USA), anti-IL-37 (eBioscience, USA), and mouse IgG (Cell Signaling Technology, USA) were used.

Techniques: Staining

Journal: Scientific Reports

Article Title: IL-37 alleviates Coxsackievirus B3-induced viral myocarditis via inhibiting NLRP3 inflammasome-mediated pyroptosis

doi: 10.1038/s41598-022-22617-y

Figure Lengend Snippet: IL-37 attenuated the activation of NLRP3 inflammasome and the production of IL-1β and IL-18. ( A ) The expression of NLRP3, ASC dimer and cleaved caspase 1 in heart tissues were measured by western blot. ( B – D ) Bar graphs of quantitative analysis of NLRP3, ASC dimer and cleaved caspase 1. ( E , F ) The levels of IL-18 and IL-1β in serum of mice in all groups were measured by ELISA. (*P < 0.01, **P < 0.001, ***P < 0.0001, ns: not significant.)

Article Snippet: Anti-IL-1R8 (Santa Cruz, USA), anti-IL-37 (eBioscience, USA), and mouse IgG (Cell Signaling Technology, USA) were used.

Techniques: Activation Assay, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: IL-37 alleviates Coxsackievirus B3-induced viral myocarditis via inhibiting NLRP3 inflammasome-mediated pyroptosis

doi: 10.1038/s41598-022-22617-y

Figure Lengend Snippet: IL-37 may decrease pyroptosis via inhibiting the expression of NF-κB. ( A ) The representative pictures of NF-κB p65 and phosphorylation of p65 expression of in heart tissues assayed by western blot. ( B , C ) Bar graphs of quantitative analysis of NF-κB p65 and phosphorylation of p65 protein. ( D ) The mRNA levels of NF-κB were detected by real time PCR. (*P < 0.01, **P < 0.001).

Article Snippet: Anti-IL-1R8 (Santa Cruz, USA), anti-IL-37 (eBioscience, USA), and mouse IgG (Cell Signaling Technology, USA) were used.

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: IL-37 alleviates Coxsackievirus B3-induced viral myocarditis via inhibiting NLRP3 inflammasome-mediated pyroptosis

doi: 10.1038/s41598-022-22617-y

Figure Lengend Snippet: Binding of IL-37 to IL-1R8 in the PBMCs. Exogenous supplementation of IL-37 enhanced the combination of IL-37 and IL-1R8. Both injection of IL-37 and CVB3 virus increase the binding of IL-37-IL-1R8 complex respectively. CVB3 infection with exogenous IL-37 enhanced the mutual binding of IL-37 and IL-1R8 (P < 0.01 VS Control and IL-37 alone). Original blots are presented in .

Article Snippet: Anti-IL-1R8 (Santa Cruz, USA), anti-IL-37 (eBioscience, USA), and mouse IgG (Cell Signaling Technology, USA) were used.

Techniques: Binding Assay, Injection, Infection

Journal: Journal of Hepatology

Article Title: Effector CD8 + T cell-derived interleukin-10 enhances acute liver immunopathology

doi: 10.1016/j.jhep.2017.04.020

Figure Lengend Snippet: IL-10 promotes liver immunopathology by preventing CD8 T E apoptosis. (A) ALT activity measured in the serum (sALT) of HBV replication-competent transgenic mice treated or not with anti-IL10Rα 2 h before the injection of 10 7 Cor93 CD8 T E . n = 5; results are representative of 3 independent experiments. (B) ALT activity measured in the serum of IL-10 +/+ or IL-10 −/− HBV replication-competent transgenic mice upon 10 7 Cor93 CD8 T E injection. n = 10; results are representative of 2 independent experiments. (C–E) Quantification of the absolute number (C), IFN-γ + (D), and apoptotic (E) Cor93 CD8 T E recovered at the indicated time points from the livers of HBV replication-competent transgenic mice that were treated or not with anti-IL-10Rα Abs prior to Cor93 CD8 T E injection. n = 5; results are representative of 2 independent experiments. (F) Histological analysis of representative HBV replication-competent transgenic mice treated or not with anti-IL10Rα 2 and sacrificed 2 days after the injection of 10 7 Cor93 CD8 T E . Arrowheads denote necroinflammatory foci. Scale bars represent 50 µm. (G) Quantitative morphometry of dead and degenerating (injured) hepatocytes and intrahepatic mononuclear and polymorphonuclear (inflammatory) cells in the same mice described in F. A minimum of 50 high power (400×) fields representing 2 mm 2 of liver tissue were examined. The results represent the mean of five observations (five mice). Results are expressed as mean + SEM.* p <0.05, *** p <0.001. Means between two groups were compared with two-tailed t test. Means among three or more groups were compared with one-way or two-way analysis of variance with Bonferroni’s post-hoc test.

Article Snippet: To assess the effect of IL-10 on Ag-induced apoptosis, Cor93 CD8 T E (10 7 cells/ml) were incubated for 1 h at 37 °C with 18 μg/ml of anti-IL-10R Ab (BioXCell), 400 ng/ml of recombinant mIL-10 (Biolegend) or left untreated prior to the addition of Cor93 peptide (1 mg/ml) and human IL-2 (1 mg/ml).

Techniques: Activity Assay, Transgenic Assay, Injection, Two Tailed Test

Journal: Journal of Hepatology

Article Title: Effector CD8 + T cell-derived interleukin-10 enhances acute liver immunopathology

doi: 10.1016/j.jhep.2017.04.020

Figure Lengend Snippet: IL-10 rescues CD8 T E from Ag-induced apoptosis by increasing IL-2 responsiveness. (A-C) Cor93 T cells were incubated with medium (white), anti-IL10Rα (18 µg/ml, light blue) or rIL-10 (400 ng/ml, dark blue). After 1 h Cor93 peptide (1 mg/ml) was added ( + Cor93) or not (Ctrl). The percentage of Annexin V+ (apoptotic) cells (A), the CD25 expression (B) and the percentage of phospo-STAT5+ cells (C) was determined on Cor93 CD8 T E 8 h, 24 h and 15 min later, respectively. n = 3; results are representative of 3 independent experiments. (D) PBMCs from 5 HLA-A201 + patients acutely infected with HBV were incubated with medium (white), anti-IL-10Rα (20 µg/ml, light blue), or rIL-10 (200 ng/ml, dark blue). After 1 h Cor18-27 peptide (1 µg/ml) was added (+Cor18) or not (Ctrl). The percentage of Annexin V+ (apoptotic) cells was determined on Core 18-27 dextramer + cells 5 h later. Results are representative of 2 independent experiments. Results are expressed as mean + SEM. * p <0.05, ** p <0.01, *** p <0.001. Means between two groups were compared with two-tailed t test. Means among three or more groups were compared with one-way or two-way analysis of variance with Bonferroni’s post-hoc test. Data in D were analyzed using Fisher’s Least Significant Difference (LSD) test.

Article Snippet: To assess the effect of IL-10 on Ag-induced apoptosis, Cor93 CD8 T E (10 7 cells/ml) were incubated for 1 h at 37 °C with 18 μg/ml of anti-IL-10R Ab (BioXCell), 400 ng/ml of recombinant mIL-10 (Biolegend) or left untreated prior to the addition of Cor93 peptide (1 mg/ml) and human IL-2 (1 mg/ml).

Techniques: Incubation, Expressing, Infection, Two Tailed Test

Journal: Biology Open

Article Title: Lateral line placodes of aquatic vertebrates are evolutionarily conserved in mammals

doi: 10.1242/bio.031815

Figure Lengend Snippet: Pharmacological inhibition of apoptosis in the posterior placodal area (PPA) of embryonic mice. (A) Summary scheme of in utero -developed control embryos including ectoderm (light grey), otic vesicle with detachment site (dark grey), epibranchial placodes (orange), and apoptosis (purple) demonstrates the peak of PPA apoptosis (compiled from , ; n =44 body sides). (B) Levels of apoptosis in the PPA (black contour in the schematized embryo; section interval evaluated=10 µm) of in utero -developed embryos ( n =20 body sides) or specimens developed for 24 h in whole embryo culture (wec). Embryos were cultured either in the presence of only the solvent DMSO (control; n =20), or in the presence of 10–100 µM of the pan-caspase inhibitor Q-VD-OPh ( n =8 for 10, 20, or 50 µM, respectively; n =20 for 100 µM), or in the presence of 200 µM of the more narrow spectrum caspase inhibitor Z-VAD-fmk ( n =12). It turned out that Q-VD-OPh treatment reduces PPA apoptosis in a dose-dependent manner. Furthermore, inhibition with 50 µM Q-VD-OPh or 200 µM Z-VAD-fmk is significantly less efficient compared with treatments using 100 µM Q-VD-OPh. Significant differences were measured with unpaired Mann–Whitney test (* P <0.001). Box plots indicate medians (centre lines), 25th and 75th percentiles (box limits), lower and upper extremes (whiskers), data points evaluated separately for each body side (purple dots), and outliers (open circles). (C) Micrographs (standardized sectioning plane) taken from anti-cleaved caspase-3 (Casp3) stained serial sections of mouse embryos treated with 10, 20, 50 or 100 µM Q-VD-OPh or 200 µM Z-VAD-fmk. It turned out that the more effective reduction of PPA apoptosis (arrowheads) is, the better rudiments of the lateral line placodes are preserved. Scale bars: 50 µm (overviews) and 10 µm (magnified insets). E, embryonic day; e1, e2, e3, epibranchial placodes 1, 2, 3, respectively; ot, otic anlage; ov, optic vesicle; p1, p2, pharyngeal pouch 1, 2, respectively.

Article Snippet: Primary antibodies included: mouse anti-Atonal homolog 1 (Atoh1; Atoh1 supernatant, Developmental Studies Hybridoma Bank, Iowa City, USA, lot 6/20/13, RRID: AB_10805299; 1:50, overnight, 4°C) ( ; ), mouse anti-β-Tubulin-III (Tubb3; clone SDL.3D10, T8660, Sigma-Aldrich, lot 073K4835, RRID: AB_477590; 1:8000, overnight, 4°C) , rabbit anti-cleaved caspase-3 (9661; Cell Signaling Technology, lot 37, RRID: AB_2341188; 1:8000, overnight, 4°C) ( , ), mouse anti-Islet 1 (Isl1; 39.4D5 ascites fluid, Developmental Studies Hybridoma Bank, lot 2/12/09, RRID: AB_2314683; 1:2000, overnight, 4°C) , goat anti-Neurogenin1 (Ngn1; sc-19231, Santa Cruz Biotechnology, lot C1215, RRID: AB_2298242; 1:100, 4 h, 37°C) , mouse anti-Neurogenin2 (Ngn2; clone 7G4, MAB3314, R&D Systems, Minneapolis, USA, lot WWI01, RRID: AB_2149520; 1:20,000, overnight, 4°C) , goat anti-Neurogenic differentiation 1 (NeuroD1; sc-1084, Santa Cruz Biotechnology, lot A0616, RRID: AB_630922; 1:800, 2 h, 37°C) , rabbit anti-Paired homeobox (Pax) 2 (71-6000; Thermo Fisher Scientific, lot 1117672A, RRID: AB_2533990; 1:4000, overnight, 4°C) , mouse anti-Pax8 (ACI 438; Biocare Medical, Pacheco, USA, lot 051712, RRID: AB_10922964; 1:100, overnight, 4°C) , rabbit anti-phospho-Histone H3 (pHH3; 9701, Cell Signaling Technology, lot 13, RRID: AB_331535; 1:1000, 2 h, 37°C) , rabbit anti-Sine oculis homeobox 1 (Six1; HPA001893, Atlas Antibodies, Stockholm, Sweden, lot A00939, RRID: AB_1079991; 1:800, overnight, 4°C) , mouse anti-Six1 (clone CLO185, AMAb90544, Atlas Antibodies, lot 02852, RRID: AB_2665581; 1:100, overnight, 4°C) that, compared to rabbit anti-Six1, was raised against the same immunogen and shows identical reactivities (present study), mouse anti-Sex determining region Y-box (Sox) 2 (clone 245610, MAB2018, R&D Systems, lots KGQ0311111, KGQ0315062, RRID: AB_358009; 1:200, overnight, 4°C) , goat anti-Sox10 (sc-17342, Santa Cruz Biotechnology, lot L2908, RRID: AB_2195374; 1:800, overnight, 4°C) , mouse anti-Sox10 (sc-365692, Santa Cruz Biotechnology, lot I0516, RRID: AB_10844002; 1:1600, overnight, 4°C) that, compared to goat anti-Sox10, produced identical staining patterns in mice (present study) and rats , and rabbit anti-T-box transcription factor (Tbx) 3 (42-4800, Thermo Fisher Scientific, lot SI256120, RRID: AB_2533526; 1:200, 2 h, 37°C) ( ).

Techniques: Inhibition, In Utero, Embryo Culture, Cell Culture, MANN-WHITNEY, Staining