Raptor and Rictor deficiency differentially affect Akt and GSK-3 phosphorylation. (A-B) Total lysates of day 12 or day 13 Cre, rap^Δ/Δ and ric^Δ/Δ BMDCs isolated using CD11c magnetic bead separation were subjected to Western blot analysis of Rictor, Raptor and p-Ser473-Akt (A) or p-Ser21/9 GSK3α/β (B) expression. Equal protein loading was confirmed by detection of actin. Bar graphs represent quantification of Raptor, Rictor, and p-Ser473-Akt (A) or p-Ser9 GSK3β (B). Expression of Raptor, Rictor, p-Ser473-Akt, and p-Ser21/9 GSK3α/β was normalized to actin and set to 100% in Cre. (A) Raptor in rap^Δ/Δ: 54.20 ± 5.68, P = .0195; Rictor in ric^Δ/Δ: 12.62 ± 2.25, P = .0027; p-Ser473-Akt in rap^Δ/Δ: 132.6 ± 0.96, P = .0084, p-Ser473-Akt in ric^Δ/Δ: 25.98 ± 5.66, P = .0073; mean ± SEM of N = 2 mice per group. Data are from one of 2 independent experiments with similar results. (B) Raptor in rap^Δ/Δ: 21.15; Rictor in ric^Δ/Δ: 8.57; p-Ser21/9 GSK3α/β in rap^Δ/Δ: 122.18, p-Ser21/9 GSK3α/β in ric^Δ/Δ: 57.32. Data shown are single values from 1 of 2 independent experiments with similar results. (C) FACS analysis of β-catenin expression in day 10 Cre (black line), rap^Δ/Δ (red line), and ric^Δ/Δ (green line) BMDCs gated on CD11c⁺ MHC-II⁺. Bar graph represents mean frequency ± SEM of N = 2 mice per group of β-catenin^high DCs gated as shown in the histogram. One of 2 independent experiments with similar results is shown.