Figure 1.
AAV FXN expressing construct and in vitro testing
(A) Structure of the AAV FXN-expressing construct. (B) Representative IF image of C2C12 cells treated with 1E+7 vg/cell AAV-hFXN or untransduced negative control of FXN (green) and nucleus (blue). Scale bars represent 100 μm. (C) Dose-response curve of FXN IF intensity to AAV-hFXN MOI (vg/cell) fit with 4PL equation to determine EC50 value. (D) Log-log plot of AAV-hFXN-driven FXN mRNA expression as a function of AAV-hFXN MOI (vg/cell). (E) Correlation plot of mRNA and protein expression following AAV-hFXN transduction. (F) Western blot probing for FXN (green) and GAPDH loading control (red), imaged on the LI-COR Odyssey system. Graphs were plotted using GraphPad Prism. IF images were taken on an Odyssey IX73 fluorescence microscope. IF, immunofluorescence; MOI, multiplicity of infection; polyA, polyadenylation signal.