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. 2018 Jan 6;9(11):9907–9924. doi: 10.18632/oncotarget.24035

Figure 8. Western Blot and RTPCR to assess protein changes in neuroblastoma cell lines treated with HCI-2509.

Figure 8

(A) Immunoblots to assess histone methylation in NGP, LAN5, and SH-SY5Y and SK-N-SH cells treated with 0, 1 and 3 µM of HCI-2509 for 48 hours. (B) P53 levels were assessed in all four cell lines treated with 0, 1 and 3 µM of HCI-2509 for 48 hours. MYCN levels were assessed in the MYCN-amplified NGP and LAN5 cells treated with 0, 1 and 3 µM of HCI-2509 for 48 hours. (C) RTPCR to assess MCYN expression in NGP, LAN5, SK-N-SH and SH-SY5Y cells upon treatment with HCI-2509 as compared to vehicle control (0). Additionally, NGP cells were pretreated with pifithrin-alpha (PFTα) and then were treated with 0 or 3 µM HCI-2509. Bars are labeled as cell line+ treatment. 0 and 3 µM represent concentrations of HCI-2509. NGP cells pretreated with PFT-α are marked. (D) Effect of LSD-1 siRNA as compared to scramble siRNA on p53, MYCN and histone methyl marks in NGP cells. (E) RTPCR demonstrating decreased transcription of LSD1 in NGP cells transfected with LSD1 siRNA as compared to scramble siRNA. Three biological replicates are represented with error bars and the relative inhibition of LSD1 expression was tested for significance by using the unpaired t-test (GraphPad Prism).