Fig. 5

Knockdown of PHGDH inhibits the translation initiation by suppressing the activities of translation initiation factors and disassembling eIF4F complex. a Western blot analysis of the phosphorylation of translation initiation related factors eIF4E, eIF4G1, eIF2α and 4E-BP1 in the constructed PANC-1 cells. b Co-immnoprecipitation results of eIF4G1, eIF4A1, PHGDH and 4E-BP1 pulled down by eIF4E in the constructed PANC-1 cells. c m⁷-GTP pull-down detection of the translation initiation complex formation in the constructed PANC-1 cells. d and e m⁷-GTP pull-down detection of the translation initiation complex formation under the treatments of 4EGI-1 and CBR5884 in PANC-1 cells. f Polysomes profiling of PANC-1-GFP and PANC-1-shPHGDH cells treated with 100 mg/ml cycloheximide for 10 min. Absorbance light at 254 nm. g Extracted the polysomes associated mRNA of PANC-1-GFP and PANC-1-shPHGDH cells and detected the ZO-1 and E-cadherin mRNA levels using qRT-PCR method. Data are representative of at least three independent experiments. p value: Student’s t-test; **p < 0.01. Columns: mean; bars: SD