FIGURE 6 | YTHDC2 regulates translation of IGF1R. (A) Verification of efficiency of FLAG-immunoprecipitation by western blot. (B) RNA immunoprecipitation of FLAG-YTHDC2 interacted with IGF1R in vivo in CNE2 cells. Protein–RNA complexes immunoprecipitated by anti-FLAG or IgG were determined by RT-PCR using specific primers for IGF1R or Hprt1 (negative control). (C) Cell lysate derived from shCON, shYTHDC2-1, and shYTHDC2-3 of CNE2-IRR cells were loaded on a linear gradient of 15–40% sucrose and centrifuged at 38,000 rpm at 4 ◦ C for 4 hr. After centrifugation, samples were then fractionated into 24 fractions (0.5mL per fraction), respectively and were monitored at 260nm. (D) The amount of IGF1R mRNA in 40S, 60S, 80S, polysome fractionated RNA and total RNA derived from shCON, shYTHDC2-1, and shYTHDC2-3 of CNE2-IRR cells were quantified by quantitative RT-PCR. Data are presented as mean ± SD from n = 3. ** P < 0.001; *** P < 0.001, student’s t-