(A) Schematic representation of wild-type PIP4K2A
locus, targeting vector, recombined allele and
FlpR/CreR-deleted null allele. Neomycin resistance
cassette flanked by Frt sites (yellow boxes), exon 2 flanked by loxP sites (red
arrows), and diphtheria toxin cassette located 3′ to exon 2. N, NdeI
restriction site.
(B) Southern blot of embryonic stem cell clones (wild-type and targeted)
using probe E3 after NdeI digestion.
(C) PCR analysis of genomic DNA derived from PIP4K2A F2
littermates.
(D) PI5P4Kα and β protein levels from brain homogenate
of PIP4K2A F2 littermates.
(E) PI5P4Kα and β protein levels in primary MEFs derived
from intercrossing PIP4K2A and PIP4K2B
knockout mice.
(F) Genotyping results of PIP4K2A/2B double
heterozygote interbreeding at weaning (observed numbers in bold, expected in
parentheses).
(G) Body weight (in grams) measurements of indicated mouse genotypes.
Results represented as the mean value ± SEM. ***p < 0.0001 with
two-tailed Student’s t test.
(H) Quantitation of PI5P levels in PIP4K2A/2B-deficient
relative to PIP4K2A/2B heterozygous MEFs. Results are from
three independent experiments and are represented as the mean value ±
SEM.
See also Figure
S6.