Figure 3.

IC₅₀ Determination and Protein Knockdown Efficacy of ASOs 05HM and 28H

(A–D) HeLa and 4T1 cells were used to generate dose-response curves by treating them with different concentrations of ASO 05HM (A and C) and 28H (B), respectively. On day 3, cell viability was determined using a CellTiter Blue Assay Kit (D). Then, cells were lysed and NPC1 and HPRT1 mRNA levels were determined. Values were normalized to the housekeeping gene HPRT1 and are shown relative to untreated cells (set as 100). IC₅₀ values were calculated using Prism 6 (GraphPad Software). Data are means and SD (triplicate wells). (E) HeLa cells were treated twice for 3 days with 10 μM ASO 05HM, 28H, negative control oligonucleotide Neg1 or were left untreated. On day 6, cells were lysed, and lysates were analyzed for NPC1 and Actin protein expression using SDS-PAGE and immunoblot analysis in duplicate wells.