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Competition binding assays. Increasing concentrations of MRS5246 or MRS5322 were added to compete for binding with [125I]I-AB-MECA to determine binding affinity, expressed as Ki or Kiapp, to recombinant mARs on HEK293 cell membranes. A & B, Both MRS5246 and MRS5322 bind to the mA3AR with much higher affinity than to the mA1AR. C, binding curves from mA3ARs to compare and show MRS5246 binds with much higher affinity than MRS5322 at the mA3AR.
