Figure 2.

(a) Schematic representation of hCDKL5 isoform_1, the selected PTCs and the peptides used to develop the commercial polyclonal and monoclonal anti-CDKL5 antibodies used in this study. (b, c) Representative western blots of full-length GFP-CDKL5 (α-CDKL5) or truncated proteins (α-GFP) showing the dose-dependent efficacy of gentamicin (Gent; b) or geneticin (G418; c). HEK293 cells were transfected with plasmids expressing the WT, R59X and R134X CDKL5 derivatives and exposed to increasing concentrations of drugs ranging from 0 to 2000 μg/ml or the corresponding vehicle (-) for 24 h. WT transfected cells were treated with 2000 μg/ml for both drugs (+). α-tubulin was used as a loading control. To avoid signal saturation, the WT loaded volume was halved. Black arrowheads indicate full-length GFP-CDKL5; white arrowheads indicate truncated proteins; empty circles indicate co-transfected GFP.