Figure 5. Ca²⁺ release from intracellular stores and tyrosine phosphorylation of PLCγ1 as stimulated by Yn-DNP₃ ligands.

a) Representative intracellular store release of Ca²⁺ in response to stimulation by the Yn-DNP₃ ligands (30 nM) as specified on individual plots. The baseline was set at 1.0, and ligand additions are indicated by arrows. Cells were suspended in BSS except that Ca²⁺ was omitted from the buffer. b) Ca²⁺ release integrated for 300 sec after ligand addition and averaged for each Yn-DNP₃ from 3 independent experiments. Values were normalized with respect to stimulation by Y16-DNP₃ for equal cell equivalents. Error bars represent standard errors. c) Representative Western blot shows tyrosine phosphorylation of PLCγ1 stimulated with the Yn-DNP₃ ligands (buffer (1), Y16-DNP₃(2), Y26-DNP₃(3), Y36-DNP₃(4), Y46-DNP₃(5)) and detected with anti-phosphotyrosine (4G10). d) The same blot was stripped and reprobed with anti-PLCγ1 to evaluate sample loading. e) Data from blots were quantified and averaged from 4 independent experiments for Yn-DNP₃ mediated tyrosine phosphorylation of PLCγ1. Values were normalized with respect to stimulation by Y36-DNP₃ for equal cell equivalents. Bars represent standard errors for the 4 experiments.