Malat1 Enhances Pro-survival Cues Downstream of T cell activation
Malat1scr/scr and WT cells containing the GP33 specific TCR transgene (P14) on the CD45.2 background were transferred separately into congenic CD45.1 WT hosts. One day later the recipient mice were infected with 2*105 p.f.u. I.p. lcmv armstrong or 2*104 c.f.u. r.o. LM-GP33. Antigen specific responses were assayed by monitoring the transferred cells by flow cytometry in the spleen.
(A-C) Bcl2 expression in transferred P14 cells in the spleen 7 and 31 days post infection for both LCMV and LM-GP33. Data from two independent experiments per LCMV time point and a single experiment per LM-GP33 time point.
(A) Representative flow cytometry plots of P14 KLRG1+ CD127- cell Bcl2 expression 7 days post infection. Numbers shown are mean fluorescence intensity.
(B) Representative flow cytometry plots of P14 CD43- CD27- cell Bcl2 expression 31 days post infection. Numbers shown are mean fluorescence intensity.
(C) Quantification of Bcl2 expression producing cells by mean fluorescence intensity within the indicated P14 subpopulation defined by KLRG1 or CD27 and CD43.
(D-E) Analysis of dead cells within splenic P14 CD43 and CD27 subpopulations at days 7 and 31 post LM-GP33 infection, data from a single experiment per time point.
(D) Representative flow cytometry plots of P14 subsets defined by CD43 and CD27. Numbers shown represent percent of dead cells per the parent subpopulation
(E) Quantification of dead cells as a percentage of parent P14 subpopulation
(F-G) Analysis of IL-2 producing P14 subsets in the spleen via IL-2 capture assay at days 7 and 31 post infection for both LCMV and LM-GP33, data from a single experiment per infection per time point.
(F) Representative flow cytometry plots of all P14 cells stained for KLRG1 and captured IL-2 from both infections at day 7. Numbers represent percent of cells in that quadrant of all P14 transferred cells.
(G) uantification of IL-2 producing cells by percent of parent within the indicated P14 subpopulation defined by KLRG1 or CD27 and CD43.
Statistics displayed determined by unpaired t-test between Malat1scr/scr and WT transferred cells, where multiple tests were performed the Holm–Šidák method was used to correct for multiple comparisons (*, p<0.05; **, p<0.01)