Figure 5.

Chloroquine addition enhances PS-ASO potency

(A) 190-HARE cells were treated with or without 0.16 μM PS-ASO (24 h) followed by 60 μM chloroquine treatment 6 h before (+6 h) or 6 h after (6 h+) PS-ASO treatment. 24 h after PS-ASO treatment, cells were assessed for malat-1 expression by qRT-PCR. Chloroquine addition alone does not affect malat-1 expression. (B) 190-HARE cells were treated with or without 60 μM chloroquine, and malat-1 expression was assessed after 24 h by qRT-PCR. Analysis of PS-ASO activity under different controls. (C) 190-HARE cells were treated with negative control Scr siRNA for 48 h, followed by with or without PS-ASO for 24 h as indicated. 6 h after PS-ASO treatment, cells received 60 μM chloroquine as indicated. 24 h after PS-ASO treatment, cells were assessed for malat-1 expression by qRT-PCR. (D) is an elaborative representation of (C) showing the effect of chloroquine in enhancing PS-ASO activity. Chloroquine acts independently of EEA1/Rab5C/Rab7A. 190-HARE cells were treated with a negative control Scr siRNA and (E) siRNA for EEA1, (F) siRNA for Rab5C, and (G) siRNA for Rab7A respectively for 48 h. PS-ASO and chloroquine treatment was done as explained for Figure 5C. Malat1 mRNA expression was quantified with qRT-PCR. Statistical analysis was performed using Student’s t test. Data presented as mean ± SEM. ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, ∗∗∗∗p ≤ 0.0001. n ≥ 3 in triplicate.