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. 2018 Aug 24;9(66):32580–32592. doi: 10.18632/oncotarget.25953

Figure 3. HELLS is a direct transcriptional target of the RB-E2F pathway.

Figure 3

(A) Chromatin immunoprecipitation (ChIP) assay reveals enrichment of E2F1 within HELLS promoter. Primers flanking the myoglobin promoter were used as negative control. Each point is mean ± s.d. of triplicate samples. (B–C) Western blot detection of HELLS protein level in osteosarcoma cells transduced with lentivirus containing shRNA against (B) E2F1 (shE2F1) show no significant change in HELLS expression, but (C) combination of E2F1 (shE2F1) and E2F3 (shE2F3) knockdown decreased HELLS protein levels. (D–E) RT-qPCR analysis of basal expression of (D) CDK4 and (E) CDK6 mRNA in different osteosarcoma cell lines normalized to mesenchymal stem cells (MSC). Each point is mean ± s.d. of triplicate samples. (F) RT-qPCR analysis of HELLS mRNA expression in osteosarcoma cell lines after Palbociclib-treated for 24 h, normalized to DMSO control cells. Each point is mean ± s.d. of triplicate samples. *p < 0.0332, ** p < 0.0021, ***p < 0.0002, ****p < 0.0001 by two-tailed t test.