![Figure 5. Loss of β-catenin suppresses elevated proliferation and apoptosis in Apc-deficient HPCs and LSKs in vivo. (A) Analysis of cell cycle status of LSKs and HPCs in BM from Mx1-Cre Apcfl/fl and Mx1-Cre Apcfl/flβ-cateninfl/fl mice 4 days after pI-pC injection. BrdU was injected into mice 24 hours before analysis (mean ± standard deviation [SD]; n = 3-4). (B) Histograms depict the mean distribution of cell cycle phases of HPCs and LSKs in BM from ApcΔ/Δβ-cateninΔ/Δ, β-cateninΔ/Δ, and β-cateninfl/fl 5 weeks after induction (mean ± SD; n = 3-4). (C) Analysis of frequency of apoptosis in HPCs and LSKs in BM from Mx1-Cre Apcfl/fl and Mx1-Cre Apcfl/flβ-cateninfl/fl mice 2 weeks after pI-pC injection (mean ± SD; n = 3-4). (D) Histograms depict the mean frequency of apoptosis of HPCs and LSKs from ApcΔ/Δβ-cateninΔ/Δ, β-cateninΔ/Δ, and β-cateninfl/fl 5 weeks after induction (mean ± SD; n = 3-4). *P < .05; **P < .01; ***P < .001. (E) Analysis of gene expression in Apc fl/fl, Apc-deficient, β-catenin–deficient, and Apc–β-catenin–deficient LSKs by quantitative PCR. The LSKs were isolated from the mice 4 days after induction. Cdkn1a, Cdkn1b, and Mcl1 are expressed at comparable levels in LSKs from β-cateninΔ/Δ and ApcΔ/Δβ-cateninΔ/Δ. Gene expression is normalized initially to actin expression. In β-catenin–deficient and Apc–β-catenin–deficient LSKs, values represent fold changes in gene expression relative to Cdkn1a in β-catenin–deficient LSKs (mean ± SD of 3 mice), whereas in Apcfl/fl and Apc-deficient LSKs, values indicate the fold changes in gene expression relative to Cdkn1a in Ap fl/fl LSKs (mean ± SD of 3-6 mice). *P < .05.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/121/20/10.1182_blood-2012-12-473470/2/4063f5.jpeg?Expires=1724237254&Signature=KwSpY2j5rRTd8GbruWKfCPE5erdXD8xALiL0lR3tSiW5W9JxnVGUNsOUrwFqXLmIXzjuP3d2cSs-heN63bf1Dn9qURoWzI9fxY4E3wqUPYy2ToHhHn6us-FduLpOUJ2pVV4L0q8nwdmlFafJ9R0kBJirtEe6ekZNWwqADWxg0T9hQSORHJdP8BzgMbsLJ32PmrABCgdD8IcCOBbKc0uRT99K2nuHJOb4kBMVN4S7VB1WlTn6qG4ML2FBoTKcU~TDE6wETXUWVgRZFou0VsppRB28p7YH4y6eFiaCt4p9iGaMWyEQ-EaXOT~cXPfEy5dIR45yawyOe1IKnWGuaLZlew__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Loss of β-catenin suppresses elevated proliferation and apoptosis in Apc-deficient HPCs and LSKs in vivo. (A) Analysis of cell cycle status of LSKs and HPCs in BM from Mx1-Cre Apcfl/fland Mx1-Cre Apcfl/flβ-cateninfl/fl mice 4 days after pI-pC injection. BrdU was injected into mice 24 hours before analysis (mean ± standard deviation [SD]; n = 3-4). (B) Histograms depict the mean distribution of cell cycle phases of HPCs and LSKs in BM from ApcΔ/Δβ-cateninΔ/Δ, β-cateninΔ/Δ, and β-cateninfl/fl 5 weeks after induction (mean ± SD; n = 3-4). (C) Analysis of frequency of apoptosis in HPCs and LSKs in BM from Mx1-Cre Apcfl/fl and Mx1-Cre Apcfl/flβ-cateninfl/fl mice 2 weeks after pI-pC injection (mean ± SD; n = 3-4). (D) Histograms depict the mean frequency of apoptosis of HPCs and LSKs from ApcΔ/Δβ-cateninΔ/Δ, β-cateninΔ/Δ, and β-cateninfl/fl 5 weeks after induction (mean ± SD; n = 3-4). *P < .05; **P < .01; ***P < .001. (E) Analysis of gene expression in Apcfl/fl, Apc-deficient, β-catenin–deficient, and Apc–β-catenin–deficient LSKs by quantitative PCR. The LSKs were isolated from the mice 4 days after induction. Cdkn1a, Cdkn1b, and Mcl1 are expressed at comparable levels in LSKs from β-cateninΔ/Δ and ApcΔ/Δβ-cateninΔ/Δ. Gene expression is normalized initially to actin expression. In β-catenin–deficient and Apc–β-catenin–deficient LSKs, values represent fold changes in gene expression relative to Cdkn1a in β-catenin–deficient LSKs (mean ± SD of 3 mice), whereas in Apcfl/fland Apc-deficient LSKs, values indicate the fold changes in gene expression relative to Cdkn1a in Apfl/fl LSKs (mean ± SD of 3-6 mice). *P < .05.
