Journal: Immunity

Article Title: T-bet transcription factor promotes antibody secreting cell differentiation by limiting the inflammatory effects of IFNγ on B cells

doi: 10.1016/j.immuni.2019.04.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: TaqMan Gene Expression Assay Ifngr2 , Applied Biosystems , Mm01210592_m1.

Techniques: Blocking Assay, Virus, Recombinant, Purification, Gene Expression, Staining, Reverse Transcription, DNA Library Preparation, cDNA Synthesis, Labeling, Microarray, Software

Journal: Cell Death Discovery

Article Title: Autophagy mediated by arginine depletion activation of the nutrient sensor GCN2 contributes to interferon- γ -induced malignant transformation of primary bovine mammary epithelial cells

doi: 10.1038/cddiscovery.2015.65

Figure Lengend Snippet: Expression of IFNGRs in cow mammary glands. Two groups of Holstein cows were fed with mixed forage (MF) or corn straw (CS). The experimental period was 12 weeks, and the pre-feeding period was 3 weeks. At the end of the feeding trial, the expression levels of IFNGR1 and IFNGR2 in mammary tissue (obtained via biopsy) were analysed using immunohistochemical staining and western blot analysis. ( a ) Immunohistochemical staining of IFNGR1 and IFNGR2. Scale bars, 100 μ m. Insets (scale bars, 200 μ m) show the overall presence of the brown colour indicating IFNGR1 and IFNGR2. Statistical analysis of the grey colour intensity (right). The data represent the mean±S.E.M. of three independent experiments. Error bars are±S.E.M. One-way ANOVA; * P <0.05. ( b ) Detection of IFNGRs via western blot analysis as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05.

Article Snippet: The main primary antibodies used in this study were specific for MAP1LC3 (Cell Signaling Technology, Beverly, MA, USA; Cat 2775), ATG5 (Novus Biologicals, Littleton, CO, USA; Cat NB110-53818), SQSTM1/p62 (Santa Cruz Biotechnology Inc., Heidelberg, Germany; Cat sc-25575), DAPDH (Cell Signaling Technology; Cat 2118), IFNGR1 (Bioss Biotechnology Co., LTD., Beijing, China; Cat bs-1463R), IFNGR2 (Bioss Biotechnology Co., LTD.; Cat bs-2710R), LC3B (Bioss Biotechnology Co., LTD; Cat bs-4843R) and SQSTM1 (Bioss Biotechnology Co., LTD; Cat bs-2951R).

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot

Journal: Cell Death Discovery

Article Title: Autophagy mediated by arginine depletion activation of the nutrient sensor GCN2 contributes to interferon- γ -induced malignant transformation of primary bovine mammary epithelial cells

doi: 10.1038/cddiscovery.2015.65

Figure Lengend Snippet: IFN- γ increased rates of autophagy in primary BMECs in vitro . ( a ) The expression of IFNGRs in BMECs. BMECs were treated with IFN- γ for 6, 24 or 48 h. At the end of the treatment period, the levels of IFNGR1 and IFNGR2 were analysed using western blot analysis with specific antibodies as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05; ** P <0.01. ( b ) BMECs were treated with IFN- γ for 3, 6, 12 or 24 h. At the end of the treatment, levels of MAP1LC3, ATG12-ATG5, SQSTM1/p62 and GAPDH were analysed using western blot analysis with specific antibodies as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05; ** P <0.01. ( c ) BMECs were treated with IFN- γ at 2.5, 5, 10 or 20 ng/ml. At the end of the treatment, levels of MAP1LC3, ATG12-ATG5, SQSTM1/p62 and GAPDH were analysed using western blot analysis with specific antibodies as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05; ** P <0.01. ( d ) Direct effect of IFN- γ on induction of autophagy. BMECs were cultured for 24 h in supernatants from either mock-treated BMECs or cells treated with 10 ng/ml of IFN- γ for 24 h in the presence or absence of anti-IFNGR1, IFNGR2 or IFNGR1/2. At the end of the treatment, levels of MAP1LC3, ATG12-ATG5, SQSTM1/p62 and GAPDH were analysed using western blot analysis with specific antibodies as described in the Materials and Methods section. The data represent the mean±S.E.M. of three independent experiments. Error bars are ±S.E.M. One-way ANOVA; * P <0.05; ** P <0.01.

Article Snippet: The main primary antibodies used in this study were specific for MAP1LC3 (Cell Signaling Technology, Beverly, MA, USA; Cat 2775), ATG5 (Novus Biologicals, Littleton, CO, USA; Cat NB110-53818), SQSTM1/p62 (Santa Cruz Biotechnology Inc., Heidelberg, Germany; Cat sc-25575), DAPDH (Cell Signaling Technology; Cat 2118), IFNGR1 (Bioss Biotechnology Co., LTD., Beijing, China; Cat bs-1463R), IFNGR2 (Bioss Biotechnology Co., LTD.; Cat bs-2710R), LC3B (Bioss Biotechnology Co., LTD; Cat bs-4843R) and SQSTM1 (Bioss Biotechnology Co., LTD; Cat bs-2951R).

Techniques: In Vitro, Expressing, Western Blot, Cell Culture