Purified Anti-Mouse TCR beta (H57-597)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Liver tissues were processed by enzymatic digestion with 2mg/mL type IV collagenase (Worthington, NY). In some experiments, cells were stimulated with 50ng/mL Phorbol 12-Myristate 13-Acetate (PMA; LC Labs, USA) and 1μg/mL Ionomycin (Enzo Life sciences, USA) for 4 hours in the presence of Golgi Stop/Golgi Plug (BD Biosciences).		Cells were analyzed using BD LSRII and FlowJo9.6.4 (TreeStar, Oregon, USA).

Upstream tips
Liver tissues were processed by enzymatic digestion with 2mg/mL type IV collagenase (Worthington, NY). In some experiments, cells were stimulated with 50ng/mL Phorbol 12-Myristate 13-Acetate (PMA; LC Labs, USA) and 1μg/mL Ionomycin (Enzo Life sciences, USA) for 4 hours in the presence of Golgi Stop/Golgi Plug (BD Biosciences).
Downstream tips
Cells were analyzed using BD LSRII and FlowJo9.6.4 (TreeStar, Oregon, USA).

Publication protocol

Liver tissues were processed by enzymatic digestion with 2mg/mL type IV collagenase (Worthington, NY). In some experiments, cells were stimulated with 50ng/mL Phorbol 12-Myristate 13-Acetate (PMA; LC Labs, USA) and 1μg/mL Ionomycin (Enzo Life sciences, USA) for 4 hours in the presence of Golgi Stop/Golgi Plug (BD Biosciences). The following antibodies were used: anti-CD3 (17A2), CD8 (53–6.7), CD4 (RM4-5), CD11c (N418), GR1 (RB6-8C5), I/A-I/E (M5/114.15.2), TCRβ (H57-597), CD45.1 (A20), IFNγ (XMG1.2) (Tonbo Biosciences, USA). Anti- Ly6C (HK1.4), anti-CD11b (M1/70), TCRγδ (GL3), goat anti-hamster IgG (Poly4055), anti-TCRVγ1.1+1.2 (4B2.9), TCRVγ2 (UC3-10A6), TCRVγ3 (536), and anti-IL-17A (TC11-18H10.1) were purchased from Biolegend (USA). Anti-mouse TCRVγ4 (49.2) and TCRVγ7 (F2.64) were the kind gift from Dr. Pablo Pereira-Esteva (Pasteur Institute, France). Cells were analyzed using BD LSRII and FlowJo9.6.4 (TreeStar, Oregon, USA).

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