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  Indian J Med Microbiol
 

Figure 5: Srcasm 178PTAP to 178AAAA/309PSAP to 309AAAA Srcasm double mutant enhances downregulation of Fyn and Src; while c-Cbl tyrosine 731 is not required for downregulating Fyn Y528F. (a) COS cells were transfected with 0.5 μg of Fyn and either 1.5 μg of Srcasm or Srcasm mutants, PTAP/PSAP and Y392F. After 16 h, cells were lysed. Lysates were subjected to SDS-PAGE and analyzed by western blot. Fyn levels decreased with Srcasm double mutant PTAP/PSAP, but not with Y392F. (b) COS cells were transfected with 0.5 μg of either Src or Src Y527F and 1.0 μg of either Srcasm or Srcasm double mutant PTAP/PSAP. After a 16-h incubation, cells were lysed. Lysates were then subjected to SDS-PAGE and analyzed by western blot. Double mutant Srcasm PTAP/PSAP downregulates Src more effectively than Srcasm while Src levels remain stable with Src Y527F. Data representative of three experiments. (c) COS cells were transfected with Fyn or FynY528F plasmids (0.5 μg) with or without plasmids expressing (2.0 μg) of c-Cbl or c-Cbl Y731F. After a 16 h incubation, cells were lysed using RIPA buffer. Lysates were then subjected to SDS-PAGE and analyzed by western blot. Data representative of three experiments

Figure 5: Srcasm <sup>178</sup>PTAP to <sup>178</sup>AAAA/<sup>309</sup>PSAP to <sup>309</sup>AAAA Srcasm double mutant enhances downregulation of Fyn and Src; while c-Cbl tyrosine 731 is not required for downregulating Fyn Y528F. (a) COS cells were transfected with 0.5 μg of Fyn and either 1.5 μg of Srcasm or Srcasm mutants, PTAP/PSAP and Y392F. After 16 h, cells were lysed. Lysates were subjected to SDS-PAGE and analyzed by western blot. Fyn levels decreased with Srcasm double mutant PTAP/PSAP, but not with Y392F. (b) COS cells were transfected with 0.5 μg of either Src or Src Y527F and 1.0 μg of either Srcasm or Srcasm double mutant PTAP/PSAP. After a 16-h incubation, cells were lysed. Lysates were then subjected to SDS-PAGE and analyzed by western blot. Double mutant Srcasm PTAP/PSAP downregulates Src more effectively than Srcasm while Src levels remain stable with Src Y527F. Data representative of three experiments. (c) COS cells were transfected with Fyn or FynY528F plasmids (0.5 μg) with or without plasmids expressing (2.0 μg) of c-Cbl or c-Cbl Y731F. After a 16 h incubation, cells were lysed using RIPA buffer. Lysates were then subjected to SDS-PAGE and analyzed by western blot. Data representative of three experiments