Figure 3.
Figure 3. Derepression of Glis2 expression in Ring1A/B-deleted MOZ-TIF2 leukemic cells. (A) Hierarchical clustering analysis of gene expression in MOZ-TIF2 leukemic cells from Ring1A−/−;Ring1Bf/f BM, Ring1A−/−;Ring1BΔ/Δ BM, Ring1A−/−;Ring1Bf/f Ink4a/Arf+/+ FL, Ring1A−/−;Ring1BΔ/Δ;Ink4a/Arf+/+ FL, Ring1A−/−;Ring1Bf/f;Ink4a/Arf−/− FL, and Ring1A−/−;Ring1BΔ/Δ;Ink4a/Arf−/− FL. 1, 2, 4, and 5 refer to results obtained from cells cultured for 48 hours in the presence or absence of 4-OHT; 3 refers to results obtained from cells cultured for 7 days. (B) Quantitative RT-PCR analysis of Glis2 expression. RNA was extracted from cells incubated with 4-OHT for 48 hours (mean ± SD, *P < .05, n = 3). (C) Western blot analysis of Ring1B, Glis2, and MOZ-TIF2 expression. Cell lysates were obtained after incubation with 4-OHT for 48 hours. (D) Immunofluorescence staining for Glis2 in MOZ-TIF2 leukemic cells incubated with 4-OHT for 96 hours. Cells were picked from vehicle-treated compact colonies or 4-OHT–treated loose colonies and attached to glass slides by cytospin. Scale bar, 50 μm. FL, fetal liver.

Derepression of Glis2 expression in Ring1A/B-deleted MOZ-TIF2 leukemic cells. (A) Hierarchical clustering analysis of gene expression in MOZ-TIF2 leukemic cells from Ring1A−/−;Ring1Bf/fBM, Ring1A−/−;Ring1BΔ/ΔBM, Ring1A−/−;Ring1Bf/fInk4a/Arf+/+ FL, Ring1A−/−;Ring1BΔ/Δ;Ink4a/Arf+/+ FL, Ring1A−/−;Ring1Bf/f;Ink4a/Arf−/− FL, and Ring1A−/−;Ring1BΔ/Δ;Ink4a/Arf−/− FL. 1, 2, 4, and 5 refer to results obtained from cells cultured for 48 hours in the presence or absence of 4-OHT; 3 refers to results obtained from cells cultured for 7 days. (B) Quantitative RT-PCR analysis of Glis2 expression. RNA was extracted from cells incubated with 4-OHT for 48 hours (mean ± SD, *P < .05, n = 3). (C) Western blot analysis of Ring1B, Glis2, and MOZ-TIF2 expression. Cell lysates were obtained after incubation with 4-OHT for 48 hours. (D) Immunofluorescence staining for Glis2 in MOZ-TIF2 leukemic cells incubated with 4-OHT for 96 hours. Cells were picked from vehicle-treated compact colonies or 4-OHT–treated loose colonies and attached to glass slides by cytospin. Scale bar, 50 μm. FL, fetal liver.

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