Figure S4.
The lipase-independent signaling role of PLCγ1.(A) Immunoblot analysis of LAT-null Jurkat T cells reconstituted with the GFP-tagged wild type or H380F PLCγ1. H380F abolishes most of the enzymatic activity of PLCγ1. Cells were stimulated with 2 µg/ml anti-CD3 and anti-CD28 antibodies for 2 min, lysed, and applied for Western blot analysis. MW, molecular weight. (B) Quantification of the level of indicated proteins, after being normalized to the expression level of GAPDH. Shown are mean ± SD; n = 3 independent experiments. Unpaired two-tailed t test was used. **, P < 0.01. (C) Cells as indicated were stimulated with 2 µg/ml anti-CD3 and anti-CD28 antibodies for 2 min, lysed, and applied for Western blot analysis. (D) The level of LAT pY171, after being normalized to the level of GAPDH, was quantified. Shown are mean ± SD; n = 3 independent experiments.

The lipase-independent signaling role of PLCγ1. (A) Immunoblot analysis of LAT-null Jurkat T cells reconstituted with the GFP-tagged wild type or H380F PLCγ1. H380F abolishes most of the enzymatic activity of PLCγ1. Cells were stimulated with 2 µg/ml anti-CD3 and anti-CD28 antibodies for 2 min, lysed, and applied for Western blot analysis. MW, molecular weight. (B) Quantification of the level of indicated proteins, after being normalized to the expression level of GAPDH. Shown are mean ± SD; n = 3 independent experiments. Unpaired two-tailed t test was used. **, P < 0.01. (C) Cells as indicated were stimulated with 2 µg/ml anti-CD3 and anti-CD28 antibodies for 2 min, lysed, and applied for Western blot analysis. (D) The level of LAT pY171, after being normalized to the level of GAPDH, was quantified. Shown are mean ± SD; n = 3 independent experiments.

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