Fig. 1

Creation of NKCC1 conditional knockout mouse line

(A) Schematics of CRISPR design and genotyping strategy. LoxP sites were inserted to flank exon 5 and exon 6 of Slc12a2 (NKCC1) gene. (B) qPCR showing reduced NKCC1 transcripts in neonatal mice following embryonic ChP NKCC1 knockdown by AAV-Cre vs. AAV-Null (empty). N = 13 AAV-Null and N = 11 AAV-Cre. ** p = 0.0062. (C) Immunoblotting of P4 ChP showing reduction in total and pNKCC1 by AAV-Cre. (D) Quantification of immunoblots. N = 4 for each condition. ** p = 0.015, **** p < 0.0001. Welch’s t-test. (E) Representative images showing robust pNKCC1 in AAV-Null mice but not in AAV-Cre mice. Scale = 100 μm