Figure 6. Phosphorylation of ELOA Ser³¹¹ by CDKL5 on damaged chromatin.

• A
  HEK293 cells were co‐transfected with CDKL5 (wild‐type “WT” or kinase‐dead “KD” K⁴²R mutant) fused to an NLS, and FLAG‐ELOA (wild‐type “WT” or a S³¹¹A mutant “SA”). Anti‐FLAG precipitates or cell extracts were probed with the antibodies indicated. One of three independent experiments is shown.
• B
  Same as (A) showing a range of pathogenic (red) and benign (blue) CDKL5 variants.
• C–E
  Wild‐type (WT), CDKL5‐disrupted (CDKL5^{Δ / Δ}) or siRNA‐transfected cells were subjected to indirect immunofluorescence analysis with the indicated antibodies at laser tracks. Quantification of ELOA‐pSer³¹¹ signal at the laser tracks is shown. Data represent mean ± SD of total pELOA Ser³¹¹ intensities in different biological replicates as indicated (n). For simplicity, only intensities greater than zero are shown. Statistical significance was assessed by one‐way ANOVA test or the unpaired t‐test with Welch's correction. Asterisks **** indicate P‐values of < 0.0001. Scale bar is 10 μm.

Source data are available online for this figure.