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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 2. MNK regulates mTORC1 signaling and DEPTOR abundance.

Figure 2

(A) HeLa cells were treated with ctrl (−), MNK1, MNK2, or MNK1/2 siRNAs (72h), harvested and analyzed by immunoblot. Quantitation represents the average of three independent series normalized by setting ctrl siRNA values to 1.

(B) Wt, MNK1, MNK2, and MNK1/2 dko MEFs were seeded into 35mm3 dishes (5 × 105 cells per dish) and harvested the next day for analysis of mTORC1-relevant proteins by immunoblot; quantitation represents the average of four independent experiments.

(C) MEFs plated as in (B) were treated with mock or IGF1 as shown, harvested, and analyzed for mTORC1 and AKT activity by immunoblot. Quantitation represents the average of two experiments normalized by setting each untreated control to 1.

HeLa (D) or HEK 293 (E) cells were treated with CGP57380 (10μM) and analyzed at the designated intervals by immunoblot. Quantitations of p-S6(SS240/4), p-4EBP1(S65) and DEPTOR represents the average of three experiments (immunoblots for a representative assay are shown in Figure S1).