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. 2019 Sep 9;47(20):10865–10880. doi: 10.1093/nar/gkz771

Figure 3.

Figure 3.

(A) Kinetics of cEt gap-mer ASO association with P54 in live cells. The LgBiT-P54nrb HeLa cell line was treated with 100 nM safe SmBiT-conjugated cEt ASO 978671 or toxic SmBiT-conjugated cEt ASO 978780. Cells were then visualized using a bioluminescent imaging microscope at 3-min intervals for a total of 2.25 h collecting brightfield and NLuc bioluminescence (blue). Arrows identify nucleolar localization of P54nrb–ASO complex. For video see Supplementary Figure S6A/B. (B) Toxic cEt gap-mer ASO association with P54nrb in live cells treated with siRNAs targeting known ASO interacting proteins. The LgBiT-P54nrb HeLa cell line was treated with an siRNAs targeting RNAse H1, P32, FUS or KPNB1 at 10 nM. Forty eight hours later cells were treated with toxic SmBiT-conjugated cEt ASO 978780 at 200 nM for 2H. Following addition of NanoGlo cells were imaged collecting substrate brightfield and NLuc. These figures are representative of numerous fields and were repeated 2–4 times. Kinetic imaging for ASO 978780 in RNAse H1 reduced cells from 0 to 2 H can be found in Supplementary Figure S7B.