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. Author manuscript; available in PMC: 2013 Sep 15.
Published in final edited form as: J Immunol. 2012 Aug 6;189(6):2824–2832. doi: 10.4049/jimmunol.1201114

Figure 1. Transient inhibition of GRK2, GRK3, GRK5 and GRK6 expression in RBL-2H3 cells and their effects in CXCR1 and CXCR2 desensitization.

Figure 1

A) RBL cells (5 × 105 cells/well) were transfected with 20 µg of control (mock) or siRNA specific for GRK2, GRK3, GRK5 or GRK6. Forty-eight h post transfection, cells were lysed and analyzed by immunoblotting. B) For receptor desensitization, GRK deficient and control (mock transfected) RBL cells (5×106 cells) expressing CXCR1 or CXCR2 were loaded with Indo-1 in the presence or absence of 10 nM CXCL8 (left and middle columns) or CXCL1 (right column) for 30 min and assayed for CXCL8 or CXCL1 (10 nM) induced intracellular Ca2+ mobilization. Traces shown are representative of 3–5 experiments. C, D and E) Desensitization were determined as percentage of control which is the peak of intracellular Ca2+ mobilization obtained in the absence of pretreatment. Data shown are average of at least 3 traces. Statistical difference is evaluated by Student’s t test. **P<0.01; *** P < 0.001.