![Fig. 5. Overexpression of 3BP2-SH2 domain suppresses FcεRI-mediated tyrosine phosphorylation of PLC-γ and calcium mobilization. / (A and B) Cells transfected with vector alone (pMT3) and cells overexpressing 3BP2-SH2 domain were stimulated with antigen for the indicated times; then cell lysates were immunoprecipitated with either anti–PLC-γ2 (A) or anti–PLC-γ1 (B) antibody. The immunoprecipitated proteins were separated by 10% SDS-PAGE and analyzed by immunoblotting with anti-pTyr mAb. The membrane was then stripped and reprobed with anti–PLC-γ2 or anti–PLC-γ1 antibody. Similar results were obtained when the other lines were examined. (C) Intracellular Ca++ concentration ([Ca++]i) was measured by means of fluorescent indicator fura-2. Sensitized cells were loaded with fura-2 and then activated with either 10 ng/mL antigen (top) or 1 μM thapsigargin (bottom). The traces show the 340:380 fluorescence ratio. Similar results were obtained when the other lines were examined.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/100/6/10.1182_blood-2001-12-0340/4/h81823126005.jpeg?Expires=1722720773&Signature=VISfHfamFRa5BAzLw8XO~DOCR4Ph7fdSH4Cu6aVClzAmh8r7AnZj43IRrn1yHxbdBbJWsH7gb0C98DYlsQgF-URmHL0Z-3BcvDAVp2mm17kYTAiyzrMDSRm5ND5P-spsigCxLJzOiZEQ1XHrOL0HbiGxT-ocQz~LusjhPcr8ZlBpmcH6SfBgqRrEhSMFESG2tq~br2Er0do-DJjIo5N00NwBLxFPaig9EPHVpnGmwbmPLdMOoK8XhVtIxXTzKkC0PKDF8p7nVEbhxJvXWJ9qlywzDybC4nOh7Vq~bVYcwgYulSiwb73aSjBpMKwU6to~1YtlM1ZCgPyDGPfdrRPAHA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Overexpression of 3BP2-SH2 domain suppresses FcεRI-mediated tyrosine phosphorylation of PLC-γ and calcium mobilization.
(A and B) Cells transfected with vector alone (pMT3) and cells overexpressing 3BP2-SH2 domain were stimulated with antigen for the indicated times; then cell lysates were immunoprecipitated with either anti–PLC-γ2 (A) or anti–PLC-γ1 (B) antibody. The immunoprecipitated proteins were separated by 10% SDS-PAGE and analyzed by immunoblotting with anti-pTyr mAb. The membrane was then stripped and reprobed with anti–PLC-γ2 or anti–PLC-γ1 antibody. Similar results were obtained when the other lines were examined. (C) Intracellular Ca++ concentration ([Ca++]i) was measured by means of fluorescent indicator fura-2. Sensitized cells were loaded with fura-2 and then activated with either 10 ng/mL antigen (top) or 1 μM thapsigargin (bottom). The traces show the 340:380 fluorescence ratio. Similar results were obtained when the other lines were examined.
