![Fig. 2. In vitro phosphorylation of FANCA. / (A) In vitro phosphorylation of Flag-wt-FANCA in GM6914 cells. Cell lysates from GM6914 and GM6914/Flag-wt-FANCA were immunoprecipitated (IP) with anti-Flag antibody (M2) or anti-FANCA antibody. For a 2-step immunoprecipitation, Flag-wt-FANCA was absorbed to anti-Flag antibody-conjugated Sepharose, eluted with Flag peptide (100 μg/mL), then immunoprecipitated with anti-FANCA antibody. Immunoprecipitates were incubated in a kinase buffer containing [γ-32P] ATP (10 μCi [0.37 MBq]) for 20 minutes. (B) A time course of in vitro phosphorylation of Flag-wt-FANCA expressed in GM6914 cells. Anti-FANCA immunoprecipitates from GM6914/Flag-wt-FANCA were incubated in a kinase buffer containing [γ-32P] ATP (10 μCi [0.37 MBq]) for the indicated times. Western blots of FANCA were shown at the bottom.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/98/13/10.1182_blood.v98.13.3650/6/h82411875002.jpeg?Expires=1724240098&Signature=Rb6SiW~y~cfqy3ltiZHaijEl~we2LJBxTwGJeoQb14Xj8VTc0ClQ5veknxUPUr6hZJ1WLCXHXiyM95vHgwr0DRIWCNskPhxRNsHzielATOoucGsyFuCJPsL1tUHyOEQOCrqDFgGVoUDsxFt0wkTWUWlwj-uLTo22v-U8upGrdPfziRFLwW5Gw0e1bTmL-GwDYbuaNDuzIQS6ba6MOoCMbSdsVpW8eLu1V4-aYoyAiOBeh6PgtPxD7nT5wOUtBv5jEONMzjCMELgswOF0bD78LuRuLxpjSyN3dGYBvJFN8uvdtiE7PHLmYRSHPck3iFNecuZqbwM2IlsHstqDPWqIzA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
In vitro phosphorylation of FANCA.
(A) In vitro phosphorylation of Flag-wt-FANCA in GM6914 cells. Cell lysates from GM6914 and GM6914/Flag-wt-FANCA were immunoprecipitated (IP) with anti-Flag antibody (M2) or anti-FANCA antibody. For a 2-step immunoprecipitation, Flag-wt-FANCA was absorbed to anti-Flag antibody-conjugated Sepharose, eluted with Flag peptide (100 μg/mL), then immunoprecipitated with anti-FANCA antibody. Immunoprecipitates were incubated in a kinase buffer containing [γ-32P] ATP (10 μCi [0.37 MBq]) for 20 minutes. (B) A time course of in vitro phosphorylation of Flag-wt-FANCA expressed in GM6914 cells. Anti-FANCA immunoprecipitates from GM6914/Flag-wt-FANCA were incubated in a kinase buffer containing [γ-32P] ATP (10 μCi [0.37 MBq]) for the indicated times. Western blots of FANCA were shown at the bottom.
