Figure 3.

ASO-005-02 can rescue TECPR2 protein expression in TECPR2-/- (SPG49) patient-derived fibroblasts

(A) Experimental outline for rescue of TECPR2 protein expression assay, using immunocytochemistry with a rabbit polyclonal antibody raised against human TECPR2 as the primary antibody. TECPR2 immunoreactivity in TECPR2-/- fibroblasts was assessed 5 days after treatment with ASO-005-02 using an IN Cell Analyzer 6000 platform. (B) Representative immunofluorescence images obtained from two fields of view (FOVs) for each of the different TECPR2 genotype and ASO treatment conditions, showing TECPR2 immunoreactivity with green signal and nuclear stain in blue. While untreated TECPR2-/- fibroblasts show no TECPR2 immunoreactivity, the same patient-derived cells treated with 100 nM or 300 nM lead ASO-005-02 show TECPR2 signal comparable with that of healthy control genotypes TECPR2+/+ and TECPR2+/- (scale bar, 20 μm). (C) Quantitative immunocytochemistry of the different TECPR2 genotype and ASO conditions. Each data point represents a single cell for which TECPR2 fluorescence signal has been estimated (n = 60–400 cells per condition, 16–32 FOVs, error bars = SEM, ∗∗∗p < 0.001 [pairwise t test FDR-corrected p value]).