Research Paper Volume 16, Issue 19 pp 12866—12892

Figure 8. FAM3D activates the mTORC1 pathway via ATF4-mediated downregulation of SESN2 and reduces the chemosensitivity of CRC cells. (A) The mRNA expression of ATF4 in FAM3D KO and NC cells. (B, C) Correlations between ATF4 mRNA expression and FAM3D mRNA expression in (B) GSE39582 and (C) GSE106582. (D, E) The mRNA and protein levels of ATF4 in LoVo cells after FAM3D (D) KO or (E) overexpression. (F) Schematic description of the luciferase reporter. (G) Relative luciferase activity of the ATF4 wild type (WT) promoter in LoVo cells with knockout or overexpression of FAM3D. (H, I) Western blot analysis of LoVo cells with (H) knockout or (I) overexpression of FAM3D. (J) Estimated IC₅₀s of oxaliplatin, cisplatin and 5-fluorouracil in FAM3D-knockout and NC cells. (K) CCK-8 assay was used to detect the viability of FAM3D-knockout or FAM3D-overexpressing LoVo cells after treatment with a series of concentrations of oxaliplatin, cisplatin, and 5-fluorouracil. *P < 0.05; **P < 0.01, ***P < 0.001, ****P < 0.0001.