Research Paper Volume 15, Issue 18 pp 9614—9632
ALDOC promotes non-small cell lung cancer through affecting MYC-mediated UBE2N transcription and regulating Wnt/β-catenin pathway
Figure 3. UBE2N is a downstream target of ALDOC. (A) The heat map of the top 20 DEGs in shALDOC-infected-A549 cells. (B) RAD51AP1, UBE2N and KIAA0101 mRNA levels were detected in A549 cells in response to ALDOC knockdown. (C) RAD51AP1 and UBE2N protein levels were detected in A549 and NCI-H1299 cells in response to ALDOC knockdown. (D) UBE2N mRNA levels were quantified in BEAS-2B, A549 and NCI-H1299 cells. (E) Immunofluorescence experiments showed the interaction between ALDOC and MYC. (F) Validation of increased MYC levels in the nuclear fraction of NSCLC cell lines after ALDOC overexpression using western blot analysis. (G) MYC and UBE2N protein expression were tested after overexpressing MYC in A549 cells. (H) Dual luciferase reporter experiment demonstrated that MYC could increase the activity of luciferase in UBE2N-WT group not in UBE2N-MUT group. (I) The transcriptional regulation of UBE2N by MYC was confirmed by ChIP-qPCR assay. *** P < 0.001.