Supplementary Materials

The PDF file includes:

  • Materials and Methods
  • Fig. S1. Assay development experiments conducted with neurons derived from ES cell lines with homozygous insertion of luciferase reporter into the KCC2 locus.
  • Fig. S2. Western blot experiments demonstrating that treatment of cultured mouse astrocytes with KEECs does not induce ectopic KCC2 expression or alter the amount of NKCC1 gene expression.
  • Fig. S3. Developmental time course of GABA reversal potential (EGABA) in cultured neonatal mouse cortical neurons.
  • Fig. S4. Pharmacological validation of GABA responses in the gramicidin-perforated patch clamp recording setup.
  • Fig. S5. Dose-response properties of KEECs to induce the KCC2 luciferase reporter signal in human RTT KCC2 reporter neurons.
  • Fig. S6. Enhancement of KCC2 gene expression in immature mouse neurons and human RTT neurons following the knockdown of the FLT3 or GSK3β gene with siRNA.
  • Fig. S7. Enhancement of the chloride extrusion rate following treatment of human RTT neurons with KW-2449.
  • Fig. S8. Increase in membrane capacitance and rescue of mEPSC amplitude deficits in human RTT neurons following KEEC treatment.
  • Fig. S9. Loss of hyperpolarizing shift in EGABA in human RTT neurons treated with KEECs following pharmacological blockade of KCC2.
  • Fig. S10. Equivalent EGABA in mature (2 to 3 months in vitro) WT human neurons treated with DMSO and those treated with KEECs.
  • Fig. S11. Equivalent frequency and median amplitude of mEPSCs in WT human neurons treated with DMSO and those treated with KEECs.
  • Fig. S12. Equivalent basic membrane properties of WT neurons treated with DMSO and those treated with KEECs and enhancement of only cell size in KEEC-treated RTT neurons.
  • Fig. S13. Representative images of neurons from different genotype and treatment groups recorded in electrophysiological experiments.
  • Fig. S14. Gramicidin-perforated patch clamp recording data presented as linear regression fits of the average GABA response recorded at different membrane holding potentials (Vhold) in different genotype and treatment groups.
  • Table S1. KEECs identified from screening with WT human KCC2 reporter neurons.
  • Table S2. KEECs identified from screening with RTT human KCC2 reporter neurons.
  • Legends for tables S3 and S4
  • References (9497)

[Download PDF]

Other Supplementary Material for this manuscript includes the following:

  • Table S3. Statistical results for all figures (provided as separate Excel file).
  • Table S4. Raw data (provided as separate Excel file).