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. Author manuscript; available in PMC: 2012 Nov 30.
Published in final edited form as: Oncogene. 2012 Feb 6;31(48):5007–5018. doi: 10.1038/onc.2012.8

Figure 1. Serine 172 phosphorylation of TAL1 controls TAL1 interaction with LSD1.

Figure 1

(A) Schematic representation of TAL1 and GST-TAL1 fusions used in the GST-pull down assay. (B) GST control and GST-TAL1 fusion proteins preabsorbed to glutathione-sepharose beads were incubated with 35S-labeled LSD1. The bound LSD1 was resolved in SDS-PAGE and visualized by fluorograpgy (Top). Coomassie blue-stained gel shows the relative GST fusion protein loading marked by asterisks (Bottom). (C) Ser172 of TAL1 is phosphorylated by PKA in vitro. The purified GST-TAL1 and its mutants were incubated with the catalytic subunit of PKA in the presence of [γ-33P] ATP, resolved in SDS-PAGE, and visualized by autoradiography. (D) The effect of Ser 172 phosphorylation on the LSD1 recruitment. The purified GST-TAL1142-185 or GST-TAL1S172A was incubated with ATP in the presence or absence of the catalytic subunit of PKA. The reaction was further incubated with purified LSD1. The bound LSD1 was resolved in SDS-PAGE and visualized by Western blotting analysis. (E) Nuclear extracts from MEL cells transduced with vector control, TAL1, TAL1S172A, and TAL1Δ142-185 were precipitated with anti-Flag conjugated beads and analyzed by Western blot with LSD1 antibody (Top) and Flag antibody (Bottom).