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. 2017 Dec 14;7:17565. doi: 10.1038/s41598-017-17770-8

Figure 5.

Figure 5

AR-12 long-term treatment cured prion infected ScN2a cells (22 L). ScN2a cells were treated with AR-12 (3 µM) or AR-14 (2 µM). DMSO treatment was used as control. Treatment was continued for five passages (20 days). Then, the treatment was stopped and cells were passaged five times again (20 days). (a,f,k and p) Immunoblots showing the effect of treatment with AR compounds on PrPSc throughout the experiment compared to DMSO treated cells. Immunoblots were developed with anti-PrP mAb 4H11 and probed for actin. (b,g,l and q) RT-QuIC analysis for uninfected N2a cells at every passage. Recombinant mouse PrP was used as substrate. Each quadruplicate RT-QuIC reaction was seeded with 2 μl of cell lysate (at dilutions 10−1 to 10−4). The average increase of Thioflavin-T fluorescence of replicate wells is plotted as a function of time. Y-axis represents relative fluorescent units (RFU) and x-axis time in hours. (c,h,m and r) RT-QuIC analysis for DMSO-treated cells. Passages 1 and 5 (P1 and P5) are shown (c,h). After discontinuation of the treatment, passages 1 and 5 (P1* and P5*) are shown (m,r). (d,i,n and s) RT-QuIC-analysis for cells treated with AR-12 (3 µM) for 20 days. Passages 1 and 5 (P1 and P5) are shown (d,i). After discontinuation of the treatment, passages 1 and 5 (P1* and P5*) are shown (n,s). (e,j,o and t) RT-QuIC analysis for cells treated with AR-14 (2 µM) for 20 days. Passages 1 and 5 (P1 and P5) are shown (e,j). After discontinuation of the treatment, passages 1 and 5 (P1* and P5*) are shown (o,t).