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Blood, Vol. 111, Issue 8, 4165-4172, April 15, 2008
PI5KI-dependent signals are critical regulators of the cytolytic secretory pathway
Blood Micucci et al.
111: 4165
Supplemental material for: Micucci et al
Files in this Data Supplement:
- Figure S1. PI5KI isoform silencing impairs CD16-induced cytotoxic in primary NK cells (PDF, 51 KB) -
Primary cultured NK cells were infected with lentiviruses encoding shRNA sequences targeting PI5KI (shRNA-ctr), PI5KI (shRNA-PI5KI) or PI5KI (shRNA-PI5KI). GFP positive cells were FACS-sorted. Total cell lysates of infected populations, Hela and PC12 cell lines were analysed by immunoblotting with the indicated Abs (top). shRNA-ctr, shRNA-PI5KI and shRNA-PI5KI populations were assessed in a 51Cr release assay against P815 target cells in the presence of anti-CD16 mAbs (bottom). In all populations the % specific lysis in the presence of control mAb (anti-MHCI) or in the absence of antibody was less than 8% at E:T ratio of 5:1. One representative experiment is shown.
- Figure S2. PI5KI isoform silencing does not impair receptor-triggered Vav tyrosine phosporylation (PDF, 33 KB) -
shRNA-ctr, shRNA-PI5KI or shRNA-PI5KI NK92 populations were stimulated with anti-2B4 mAb for 5 min. Vav tyrosine phosphorylation status was evaluated by immunoblot analysis with anti-pTyr mAb. The same membranes were reprobed with anti-Vav mAb for sample normalization. One representative experiment is shown.
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