Supplementary Fig. 1.

A: Sub-cellular localization of CDKL5. Representative fluorescence image of SH-SY5Y cells expressing CDKL5-FLAG. Twenty-four hours after transfection, SH-SY5Y cells were immunostained for FLAG (red signal) and Hoechst dye (blue signal). Cells show both CDKL5 nuclear and cytoplasmic localization. Scale bar: 40 μm. B: FLAG-tagged CDKL5 was transiently over-expressed in SH-SY5Y cells and detected in nuclear and cytoplasmatic extracts 24 h after transfection. Example of Western blot with anti-FLAG antibodies. Histon H1 (H1) and β-actina (β-ACT) were used respectively as nuclear and cytoplasmic markers. C: FLAG-tagged CDKL5 was transiently over-expressed in SH-SY5Y cells and detected 24, 48 and 72 h after transfection. Total cell extracts were analyzed by Western blotting with anti-FLAG and β-actina antibodies.