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. 2020 Nov 12;6(11):e05442. doi: 10.1016/j.heliyon.2020.e05442

Figure 5.

Figure 5

Silencing ALDH1A1 and NEK-2 in A2780 cells and its impact on ABC membrane transporters. A) NEK-2 expression in A2780 and A2780-Cp ovarian cancer cells. Relative expression for NEK-2 mRNA with respect to the gene GAPDH presented as fold change. All samples were run in triplicate and relative gene expressions were determined by 2-ΔΔCt method with SYBR-green real-time PCR. Data were expressed as Mean ± SD. ∗P < 0.05 and ∗∗P < 0.01 between control and siRNA treated cells. B) Effect of ALDH1A1 silencing on NEK-2 expression in A2780 and A2780-Cp cells. C) Effect of ALDH1A1 silencing on ABC transporters expression (ABCB1, ABCG2 and ABCC1) in A2780-Cp cells. D) Effect of NEK-2 silencing on ABC transporter expression (ABCB1, ABCG2 and ABCC1) in A2780-Cp cells. E) Impact of ALDH1A1 and NEK-2 silencing on downstream molecules was assessed by western blotting. Bar diagram showing the protein expression compared to GAPDH determined by densitometric analysis of proteins bands. F) Overexpression of NEK-2 in A2780 cells, its confirmation at mRNA and protein levels and determination of ABCB1 expression. Bar diagram showing the protein expression compared to GAPDH determined by densitometric analysis of proteins bands.