Fig. 6: LNA-flanked amides and PS linkages increase gymnotic delivery, exon-skipping activity and reduce toxicity.

In all cases, luciferase activity was measured and normalised to both protein quantity and untreated cells. Experimental details and sequences are given in the Supplementary Information. a HeLa pLuc/705 cells were transfected at 100 nM ON using LF2000. b Dose-response for active ONs in a. c Cells were treated with ONs without a transfection agent. d Viability of the HeLa pLuc/705 cells following lipofection with ONs using LF2000 determined using WST-1 assay. For all graphs (a–d) data were means ± SD and values for ON18^{2′OMe/4LAL/13PS} and ON20^2′OMe/17PS were compared using unpaired two-sided t-test analysis. ns represents P > 0.05, *represents P < 0.05, **represents P < 0.01, ***represents P < 0.001. Each dot represents one distinct replicate (n = 3). ON31^{2′OMe/17PS scrambled}: CCUCAUUCACUCGAUUCA. e Change in cell viability after treatment with 200 nM ONs complexed with LF2000 compared to untreated, scale bar = 200 μm. Micrographs of cells shown are the representations from ≥10 images taken per condition. f Reversed-phase HPLC chromatogram and g. Electrospray (ES-) mass spectrum of purified ON18 used in the assays (required mass = 5976.2, found mass = 5975.5). Source data are provided in the Source Data file.