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. Author manuscript; available in PMC: 2017 Feb 23.
Published in final edited form as: Cell Rep. 2017 Feb 7;18(6):1444–1457. doi: 10.1016/j.celrep.2017.01.023

Figure 5. MNK regulates the association of TELO2:DDB1 with a complex containing MNK, mTOR, and Raptor.

Figure 5

(A, B) Flag-TELO2 (A) or Flag-DDB1 (B) were transfected in HEK293 cells pretreated with DMSO or CGP57380 as shown. Lysates were subjected to Flag-IP and immunoblot.

(C) HEK293 cells were transfected with empty vector (PC3), Flag-MNK1, or Flag-MNK2 and treated with DMSO, TPA, or TPA + CGP57380 as shown. Flag-IP of lysates and immunoblot analysis was performed to determine changes in MNK associations defined in Figure 4.

(D) HEK293 cells were transfected with ctrl or TELO2-targeting siRNA followed by transfection with empty vector (PC3) or Flag-MNK2. MNK2 IP and immunoblot analysis were performed as in (C). The assay was performed 3 times; a representative series is shown.

(E) Dox-inducible HA-Raptor expressing HEK293 cells were treated with mock (DMSO) or TPA, lysates were prepared and subjected to HA-IP, and immunoprecipitated proteins were analyzed by immunoblot as shown.

(A–E) Quantitations are averages from 3 (A, C, E) or 2 (B) independent assays, normalizing control values to 1.