Figure 2

Transfection of cystinosis patient (CYS) derived proximal tubular epithelial cells (PTECs) and podocytes (PODOs) results in functional cystinosin-3HA expression for up to 4 and 10 days, and reduces cystine levels for up to 10 and 18 days, respectively. (a,b) Time course of cystinosin-3HA expression in PTECs (a) and PODOs (b). Images were obtained using the Operetta CLS High Content Screening Microscope and percentage of cells with detectable cystinosin-3HA expression quantified (each dot represents one observed field from 3 independent experiments). Data were analysed using a Kruskal Wallis test (PTECs) and Welch’s ANOVA test (PODOs). Median and 95% CI are represented for PTECs, and mean and SEM are shown for the PODOs. *p < 0.05; ***p < 0.001; ****p < 0.0001. (c,d) Time course of cystine reduction in CYS PTECs (c) and PODOs (d) that were transfected with CTNS-3HA mRNA. Cystine levels were measured at 4, 10, 14 and 18 days post-transfection and were expressed as percentage (%) of levels measured in cells transfected with GFP mRNA. Each dot represents a replicate, coming from 2 independent transfections. Statistical significance was evaluated using individual tests for each time point, one-way ANOVA (****p < 0.001) and Kruskal Wallis test (^$$p < 0.01; ^$$$p < 0.001; ^$$$$p < 0.0001). Median and 95% CI are indicated for all time points.