Journal: Cell Reports

Article Title: Stromal Cell-Contact Dependent PI3K and APRIL Induced NF-κB Signaling Prevent Mitochondrial- and ER Stress Induced Death of Memory Plasma Cells

doi: 10.1016/j.celrep.2020.107982

Figure Lengend Snippet:

Article Snippet: Anti-mouse BCL2, APC, REA356 , Miltenyi Biotec , Catalog # 130-105-474; RRID: AB_2651266.

Techniques: Recombinant, Electron Microscopy, Staining, Software

Journal: International immunology

Article Title: Effects of BCL-2 over-expression on B cells in transgenic rats and rat hybridomas.

doi: 10.1093/intimm/dxr071

Figure Lengend Snippet: Fig. 6. Hybridoma characterization. (A) Binding constants (nanomolar) of mAbs generated with splenocytes from non-transgenic littermates (LM) and transgenic line 70818 animals (Bcl2) as determined by Biacore analysis (log scale nanomolar). (B) Western blot analysis of Bcl-2 expression in hybridomas from bcl-2 transgenic animals (each line represents one hybridoma). The positive control corresponds to a lysate from human Jurkat cells. The ratio of Bcl-2/GAPDH for each hybridoma is depicted under the respective lines. (C) Concentration of rat IgG in tissue culture supernatants at day 3 of culture (cells between 3.1 and 6 3 105 ml–1) measured by ELISA. Each point represents the values of one hybridoma supernatant and the horizontal bar the mean of each group of values *P < 0.05 hybridoma Bcl-2- (n = 4) versus hybridoma Bcl-2+

Article Snippet: Western blots were performed as previously described (26) using a mouse anti-human Bcl2 mAb (Abd Serotec) followed by incubation with HRP-conjugated goat anti-mouse IgG + IgM (H + L) antibodies (Jackson ImmunoResearch).

Techniques: Binding Assay, Generated, Transgenic Assay, Western Blot, Expressing, Positive Control, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell

Article Title: Long-term programming of CD8 T cell immunity by perinatal exposure to glucocorticoids

doi: 10.1016/j.cell.2020.02.018

Figure Lengend Snippet: (A and B) Footpads of 13 week old WT (Nr3c1fl/fl) mice and KO (CD4-Cre Nr3c1fl/fl) mice were immunized and analyzed (described in Figure 1B). Flow cytometric analysis of dead CD8 T cells in LNs (n=6–8/group (female and male)) (A), and the expression of Bcl2 and CD69 in CD8 T cell (n=6–8/group (female and male)) (B). (C and D) Wild-type BMDC and naïve CD8 T cells from 14 week old WT and KO mice were co-cultured in the presence or absence of anti-CD3ε antibody and IL-12. Flow cytometric analysis of CD25+ IFN-γ+ CD8 T cells (n=3/group (female), representative of 3 independent experiments including males) (C). Flow cytometric analysis of expression of proteins in CD8 T cells after activation with anti-CD3ε antibody (n=6/group (female), representative of 2 independent experiments) (D). (E-G) CD8 T cells were sorted (single cells/live/CD45+/CD3ε+/CD8α+) from LNs of 12 week old male B6 mice after 5 days of footpad immunization. mRNA from CD8 T cells was isolated and analyzed with RNA-seq. Differentially-regulated top canonical pathways with IPA analysis (E). Representative transcripts per millions (TPM) (n=3/group) (F and G). (H) Wild-type BMDC and naïve OT-I cells sorted from 16 week old OT-I mice were co-cultured as described in Figure 1G. Flow cytometric analysis after 3–6 hours of SIINFEKL (OVAp) treatment with or without IL-12 (n=3/group (male), representative of 2 independent experiments including females). Data are represented as mean ± SEM. 2-Way ANOVA (C and H), Student’s t-test (rest), *p<0.05, **p<0.01, ***p<0.001 vs Control or wild-type (WT) group.

Article Snippet: Anti-mouse Bcl2 APC (Clone REA356) , Miltenyi Biotec , Cat#130-105-431;RRID: AB_2651267.

Techniques: Expressing, Cell Culture, Activation Assay, Isolation, RNA Sequencing, Control

Journal: Cell

Article Title: Long-term programming of CD8 T cell immunity by perinatal exposure to glucocorticoids

doi: 10.1016/j.cell.2020.02.018

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-mouse Bcl2 APC (Clone REA356) , Miltenyi Biotec , Cat#130-105-431;RRID: AB_2651267.

Techniques: Purification, Virus, Expressing, Recombinant, Adjuvant, Enzyme-linked Immunosorbent Assay, Staining, Isolation, Cell Isolation, Software