Transcriptional regulation and PI3K-Akt/Notch pathways after PEAR1 knockdown. (A) Proportional mRNA expression of the indicated transcription factors on day 11 in CD34⁺ cells transduced with control or LV_miR_PEAR1_1461 vectors vs GAPDH. (B) List of genes involved in the Notch and PI3K/Akt pathways under- or overexpressed in RT²  profiler analysis in the shPEAR1 knockdown, expressed in fold change. (C) qRT-PCR for PTEN mRNA in CD34⁺ cells 10 days after transduction with control and LV_miR_PEAR1_1461, relative to GAPDH; western blot for PTEN at day 10 after transduction with control and LV_miR_PEAR1_2938, relative to GAPDH in 2 separate analyses (A-B). (D) qRT-PCR for HES1 mRNA in CD34⁺ cells at day 10 after transduction with control and LV_miR_PEAR1_1461, relative to GAPDH. (E) Western blots for AKT-P, Akt, ERK1/2-P, and ERK1/2 in control and LV_miR_PEAR1_1461 treated CD34⁺ cells at day 10. Phosphorylation was quantified with ImageJ and is shown in the right histogram.