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. Author manuscript; available in PMC: 2014 Jun 25.
Published in final edited form as: Cell. 2013 Nov 7;155(4):844–857. doi: 10.1016/j.cell.2013.09.057

Figure 6. Requirement for PIP4K2A and PIP4K2B for Survival and Growth.

Figure 6

(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.