Incubation:Article Title: Development and Internal Validation of a Novel Pre-Transplant Biomarker Panel to Predict Post-Liver Transplant Mortality
Article Snippet: .. Viability dye Ghost Dye Violet 510 (Tonbo Biosciences, San Diego, CA) was added, followed by incubation for 5 minutes at room temperature. .. Antibodies for cell surface staining were then added containing the following: anti-Human CD3 mAb (Brilliant Violet 605, UCHT1), anti-Human CD4 mAb (PerCP-Cy5.5, RPA-T4), anti-Human CD8a mAb (APC-Cy7, RPA-T8), antiHuman CCR7 mAb (Brilliant Violet 711, G043H7), anti-Human CD45RA mAb (FITC, HI100), antiHuman PD-1 mAb (PE-Cy7, EH12.2H7), anti-Human Tim-3 mAb (CD366, Brilliant Violet 421, F382E2), anti-Human TIGIT mAb (PE, A15153G) (all BioLegend, San Diego, CA), and antiHuman CD25 mAb (BUV395, 206D, BD Biosciences, Franklin Lakes, NJ).
Article Title: Mitochondrial ATP production promotes T cell differentiation and function by regulating chromatin accessibility
Article Snippet: .. T cells were treated with Cell Activation Cocktail for 1 h, followed by Brefeldin A (both BioLegend) treatment for 3 h. Cells were harvested, washed with cold PBS and incubated for 10 min with a solution containing Human TruStain FcX Fc Receptor Blocking Solution (BioLegend) and the viability dye Ghost Dye Violet 510 (Cytek). .. Thereafter, cells were washed with FACS buffer (2% FBS in PBS) and stained for 45 min at 4 °C with antibodies targeting surface markers using BD Horizon Brilliant Stain Buffer (BD Biosciences).
Staining:Article Title: Limited Impact of the Inhibitory Receptor TIGIT on NK and T Cell Responses during Toxoplasma gondii Infection.
Article Snippet: .. Cells were stained with the viability dye Ghost Dye Violet 510 (12- 0870; Tonbo Biosciences), and the following Abs were used for subsequent staining: IFN-g FITC (XMG1.2; Invitrogen), CD11c https://doi.org/10.4049/immunohorizons.2100007 D ow nloaded from http://journals.aai.org/im m unohorizons/article-pdf/5/6/384/1485649/ih2100007.pdf by guest on 14 O ctober 2024 FITC (N418; eBioscience), CD11a PerCP-Cy5.5 (M17/4; BioLegend), Eomes eFluor 450 (Dan11mag; Invitrogen), F4/80 eFluor 450 (BM8; Invitrogen), CD19 Brilliant Violet 605 (6D5; Bio Legend), CD4 Brilliant Violet 650 (RM4-5; BioLegend), CD11b Brilliant Violet 650 (m1/70; BioLegend), XCR1 Brilliant Violet 650 (ZET; BioLegend), CD19 Brilliant Violet 650 (6D5; Bio Legend), CD11b Brilliant Violet 650 (m1/70; BioLegend), CD155 Brilliant Violet 711 (TX56; BioLegend), CD3 Brilliant Violet 785 (17A2; BioLegend), TIGIT eFluor 660 (GIGD7; Invitrogen), CD4 AF700 (RM4-5; BioLegend,), MHC class II (MHC-II) AF700 (M5/114.15.2; BioLegend), NK1.1 allophycocyanin-Cy7 (PK136; BD Biosciences), CD45 allophycocyanin-eFluor780 (30- F11; Invitrogen), CD4 allophycocyanin-eFluor780 (RM4-5; Invitrogen), NK1.1 PE (PK136; BioLegend), CD8 PE-CF594 (53-; BioLegend), CD226 (10E5; BioLegend). ..
Article Title: IL-27 limits HSPC differentiation during infection and protects from stem cell exhaustion
Article Snippet: .. Cells were stained with the viability dye Ghost Dye Violet 510 (Tonbo biosciences; 12-0870), and the following antibodies were used for subsequent staining: CD90.2 (BD; clone 30-H12), B220 (BD; clone RA3-6B2), Ly6G (BD; clone 1A8), CD11b (BD; clone M1/70), Ly6C (Biolegend; clone HK1.4), Sca-1 (BD/Invitrogen; clone D7), CD117/cKit (Biolegend; clone ACK2), CD135 (Biolegend; clone A2F10), CD150 (Biolegend; clone TC15-12F12.2), CD48 (Biolegend/BD; clone HM48-1), CD127 (Biolegend; clone A7R34), CD16/32 (BD; clone 2.4G2), CD34 (Invitrogen; clone RAM34), CD115 (Biolegend; clone AFS98), CCR2 (Biolegend; clone SA203G11), CX3CR1 (Biolegend; clone SA011F11), CD11c (Biolegend; clone N418), XCR1 (Biolegend; clone ZET), CD172a (Biolegend; clone P84), CD64 (Biolegend; clone X54-5/7.1), F4/80 (Invitrogen; clone BM8), MHCII (Biolegend; clone M5/114.15.2), gp130 (Biolegend; clone 4H1B35), IL-27R⍺ (BD; 2918), CD132 (BD; clone 4G3), CD131 (BD; clone JORO50), IL-6R⍺ (Biolegend; clone D7715A7), GM-CSFR⍺ (R&D systems; clone 698423), CD210/IL-10R (Biolegend; clone 1B1.3a), IFN-γR β (Biolegend; clone MOB-47), IFNAR-1 (Biolegend; clone MAR1-5A3), IL-21R (Biolegend; clone 4A9), CD120a/TNFRI (Biolegend; clone 55R-286), CD120b/TNFRII (Biolegend; clone TR75-89), and CD217/IL-17RA (Biolegend; clone S19010F). .. Intracellular staining for Ki67 (BD; clone B56), and BCL2 (Biolegend; clone BCL/10C4) was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience) according to the manufacturer’s instructions.
Article Title: Single-cell analysis identifies the interaction of altered renal tubules with basophils orchestrating kidney fibrosis.
Article Snippet: 1Department of Medicine, Renal Electrolyte and Hypertension Division, University of Pennsylvania, Philadelphia, PA, USA.. 2Department of Pathobiology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA, USA.. 3Center for Immunity and Inflammation, Department of Medicine, New Jersey Medical School, Rutgers–The State University of New Jersey, Newark, NJ, USA.
Article Title: IL-27 limits HSPC differentiation during infection and protects from stem cell exhaustion
Article Snippet: .. Cells were stained with the viability dye Ghost Dye Violet 510 (Tonbo Biosciences; 12- 0870), and the following antibodies were used for subsequent staining: CD90.2 (BD; clone 30- H12), B220 (BD; clone RA3- 6B2), Ly6G (BD; clone 1A8), CD11b (BD; clone M1/70), Ly6C (Biolegend; clone HK1.4), Sca- 1 (BD/Invitrogen; clone D7), CD117/cKit (Biolegend; clone ACK2), CD135 (Biolegend; clone A2F10), CD150 (Biolegend; clone TC15- 12F12.2), CD48 (Biolegend/BD; clone HM48- 1), CD127 (Biolegend; clone A7R34), CD16/32 (BD; clone 2.4G2), CD34 (Invitrogen; clone RAM34), CD115 (Biolegend; clone AFS98), CCR2 (Biolegend; clone SA203G11), CX3CR1 (Biolegend; clone SA011F11), CD11c (Biolegend; clone N418), XCR1 (Biolegend; clone ZET), CD172a (Biolegend; clone P84), CD64 (Biolegend; clone X54- 5/7.1), F4/80 (Invitrogen; clone BM8), MHCII (Biolegend; clone M5/114.15.2), gp130 (Biolegend; clone 4H1B35), IL- 27R⍺ (BD; 2918), CD132 (BD; clone 4G3), CD131 (BD; clone JORO50), IL- 6R⍺ (Biolegend; clone D7715A7), GM- CSFR⍺ (R&D Systems; clone 698423), CD210/IL- 10R (Biolegend; clone 1B1.3a), IFN-γR β (Biolegend; clone MOB- 47), IFNAR- 1 (Biolegend; clone MAR1- 5A3), IL- 21R (Biolegend; clone 4A9), CD120a/TNFRI (Biolegend; clone 55R- 286), CD120b/TNFRII (Biolegend; clone TR75- 89), and CD217/ IL- 17RA (Biolegend; clone S19010F). .. Intracellular staining for Ki67 (BD; clone B56), and BCL2 (Biolegend; clone BCL/10C4) was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience) according to the manufacturer’s instructions.
Article Title: IL-27 limits HSPC differentiation during infection and protects from stem cell exhaustion
Article Snippet: .. Cells were stained with the viability dye Ghost Dye Violet 510 (Tonbo Biosciences; 12-0870), and the following antibodies were used for subsequent staining: CD90.2 (BD; clone 30-H12), B220 (BD; clone RA3-6B2), Ly6G (BD; clone 1A8), CD11b (BD; clone M1/70), Ly6C (Biolegend; clone HK1.4), Sca-1 (BD/Invitrogen; clone D7), CD117/cKit (Biolegend; clone ACK2), CD135 (Biolegend; clone A2F10), CD150 (Biolegend; clone TC15-12F12.2), CD48 (Biolegend/BD; clone HM48-1), CD127 (Biolegend; clone A7R34), CD16/32 (BD; clone 2.4G2), CD34 (Invitrogen; clone RAM34), CD115 (Biolegend; clone AFS98), CCR2 (Biolegend; clone SA203G11), CX3CR1 (Biolegend; clone SA011F11), CD11c (Biolegend; clone N418), XCR1 (Biolegend; clone ZET), CD172a (Biolegend; clone P84), CD64 (Biolegend; clone X54-5/7.1), F4/80 (Invitrogen; clone BM8), MHCII (Biolegend; clone M5/114.15.2), gp130 (Biolegend; clone 4H1B35), IL-27R⍺ (BD; 2918), CD132 (BD; clone 4G3), CD131 (BD; clone JORO50), IL-6R⍺ (Biolegend; clone D7715A7), GM-CSFR⍺ (R&D Systems; clone 698423), CD210/IL-10R (Biolegend; clone 1B1.3a), IFN-γR β (Biolegend; clone MOB-47), IFNAR-1 (Biolegend; clone MAR1-5A3), IL-21R (Biolegend; clone 4A9), CD120a/TNFRI (Biolegend; clone 55R-286), CD120b/TNFRII (Biolegend; clone TR75-89), and CD217/IL-17RA (Biolegend; clone S19010F). .. Intracellular staining for Ki67 (BD; clone B56), and BCL2 (Biolegend; clone BCL/10C4) was performed using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience) according to the manufacturer’s instructions.
Immunopeptidomics:Article Title: Limited Impact of the Inhibitory Receptor TIGIT on NK and T Cell Responses during Toxoplasma gondii Infection.
Article Snippet: .. Cells were stained with the viability dye Ghost Dye Violet 510 (12- 0870; Tonbo Biosciences), and the following Abs were used for subsequent staining: IFN-g FITC (XMG1.2; Invitrogen), CD11c https://doi.org/10.4049/immunohorizons.2100007 D ow nloaded from http://journals.aai.org/im m unohorizons/article-pdf/5/6/384/1485649/ih2100007.pdf by guest on 14 O ctober 2024 FITC (N418; eBioscience), CD11a PerCP-Cy5.5 (M17/4; BioLegend), Eomes eFluor 450 (Dan11mag; Invitrogen), F4/80 eFluor 450 (BM8; Invitrogen), CD19 Brilliant Violet 605 (6D5; Bio Legend), CD4 Brilliant Violet 650 (RM4-5; BioLegend), CD11b Brilliant Violet 650 (m1/70; BioLegend), XCR1 Brilliant Violet 650 (ZET; BioLegend), CD19 Brilliant Violet 650 (6D5; Bio Legend), CD11b Brilliant Violet 650 (m1/70; BioLegend), CD155 Brilliant Violet 711 (TX56; BioLegend), CD3 Brilliant Violet 785 (17A2; BioLegend), TIGIT eFluor 660 (GIGD7; Invitrogen), CD4 AF700 (RM4-5; BioLegend,), MHC class II (MHC-II) AF700 (M5/114.15.2; BioLegend), NK1.1 allophycocyanin-Cy7 (PK136; BD Biosciences), CD45 allophycocyanin-eFluor780 (30- F11; Invitrogen), CD4 allophycocyanin-eFluor780 (RM4-5; Invitrogen), NK1.1 PE (PK136; BioLegend), CD8 PE-CF594 (53-; BioLegend), CD226 (10E5; BioLegend). ..
Activation Assay:Article Title: Mitochondrial ATP production promotes T cell differentiation and function by regulating chromatin accessibility
Article Snippet: .. T cells were treated with Cell Activation Cocktail for 1 h, followed by Brefeldin A (both BioLegend) treatment for 3 h. Cells were harvested, washed with cold PBS and incubated for 10 min with a solution containing Human TruStain FcX Fc Receptor Blocking Solution (BioLegend) and the viability dye Ghost Dye Violet 510 (Cytek). .. Thereafter, cells were washed with FACS buffer (2% FBS in PBS) and stained for 45 min at 4 °C with antibodies targeting surface markers using BD Horizon Brilliant Stain Buffer (BD Biosciences).
Blocking Assay:Article Title: Mitochondrial ATP production promotes T cell differentiation and function by regulating chromatin accessibility
Article Snippet: .. T cells were treated with Cell Activation Cocktail for 1 h, followed by Brefeldin A (both BioLegend) treatment for 3 h. Cells were harvested, washed with cold PBS and incubated for 10 min with a solution containing Human TruStain FcX Fc Receptor Blocking Solution (BioLegend) and the viability dye Ghost Dye Violet 510 (Cytek). .. Thereafter, cells were washed with FACS buffer (2% FBS in PBS) and stained for 45 min at 4 °C with antibodies targeting surface markers using BD Horizon Brilliant Stain Buffer (BD Biosciences).
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