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. 2002 Jul;22(13):4535–4543. doi: 10.1128/MCB.22.13.4535-4543.2002

FIG. 1.

FIG. 1.

hnRNP K, but not hnRNP E1, is a substrate and activator of c-Src. HeLa cells were transiently transfected with His-tagged hnRNP K (A to G) or FLAG-tagged hnRNP E1 (H to N) and c-Src, the activated mutants Src(KP) and Src(Y527F), or the inactive autophosphorylation site mutant Src(Y416F). HeLa cell lysate was resolved by SDS-PAGE and analyzed in Western blot assays (lanes 1 to 8) with either anti-His (A), anti-hnRNP E1 (H), anti-c-Src (B and I), antiphosphotyrosine (p-Tyr) (C and J), or anti-Src(416) (D and K) antibodies. His-tagged hnRNP K was immunoprecipitated with an anti-His antibody (E to G) and FLAG-tagged hnRNP E1 with an anti-FLAG antibody (L to N), resolved by SDS-PAGE, and analyzed by Western blot assays with antibodies against the His tag (E) or the FLAG tag (L) to assess the amounts of precipitated hnRNP K and hnRNP E1, p-Tyr to analyze the Tyr phosphorylation status of hnRNP K and hnRNP E1 (F and M), or Src antibody to visualize coprecipitated c-Src (G and N). The increased background in panels H to N results from the necessity of exposing the Western blots significantly longer than those in panels A to G to discern the specific signals. wt, wild type.