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. 2015 Jul 27;212(8):1219–1237. doi: 10.1084/jem.20141254

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© 2015 Kang et al.

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Figure 4. — PLD1 inactivation down-regulates expression of β-catenin. (A–D) IHC for β-catenin in tumor tissues (A) and normal mucosa (B) of Apc^Min/+ and Apc^Min/+Pld1^−/− mice and in tumor tissues (C) and normal mucosa (D) of Apc^Min/+/vehicle and Apc^Min/+/PLD1-Inh mice. The nuclear β-catenin in the tumor was quantified (A and C, right). n = 21 per group; five tumors per mouse. (E) IHC for PLD1 (top) and β-catenin (bottom) in normal colon mucosa of non-AOM/DSS mice (n = 4) and tumor tissues of AOM/DSS-Pld1^+/+ and AOM/DSS-Pld1^−/− mice. The nuclear β-catenin in the tumor was quantified (right). n = 9 per group; four tumors per mouse. (A–E) Results are representative of at least three independent experiments and are shown as mean ± SEM. A Chi-square test was used. (F) q-RT-PCR analysis of the indicated mRNAs in tumor tissues of Apc^Min/+ and Apc^Min/+Pld1^−/− mice (left) or Apc^Min/+/vehicle and Apc^Min/+/PLD1-Inh mice (right). (G) q-RT-PCR analysis of the indicated mRNAs in tumor tissues of AOM/DSS-Pld1^+/+ and AOM/DSS-Pld1^−/− mice (left) or vehicle- and PLD1 inhibitor–treated AOM/DSS mice (right). (F and G) Results are representative of at least two independent experiments and are shown as mean ± SEM. n = 8 per group. A Student’s t test was used. (H) q-RT-PCR analysis of β-catenin in PLD1-depleted stable cells. Results are representative of three independent experiments and are shown as mean ± SEM. A Student’s t test was used. (I) Effect of 10 µM PLD1 inhibitor on the protein stabilization of β-catenin in cells treated with 50 µM cycloheximide (CHX) for the indicated times. Results are representative of at least three independent experiments. (J) β-catenin mRNA expression is inversely correlated with survival in 174 CRC patient specimens. Survival probability was analyzed using Kaplan-Meier, and differences were evaluated using the log-rank test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. MW, molecular weight; a.u., arbitrary units. Bars, 100 µm.