Research Paper Volume 12, Issue 9 pp 8067—8083

BMAL1 knockdown triggers different colon carcinoma cell fates by altering the delicate equilibrium between AKT/mTOR and P53/P21 pathways

Figure 1. Lentiviral ShBMAL1 decreased BMAL1 expression in three CRC cell lines but only altered expression of some circadian genes in primary CRC cell lines. (A) Effect of shBMAL1 on BMAL1 mRNA level was ascertained by quantitative RT-PCR. 36B4 was used as a quantitative reference (n=5; *p<0.05; ***p<0.001; ****p<0.0001). (B) Effect of shBMAL1 on the level of BMAL1 protein was ascertained by Western-blot. Left, a representative immunoblot is shown. Right, Bar charts represent BMAL1 expression normalized to HSC70 (n=3; *p<0.05). Data are shown as mean ± SEM. (C) Quantitative RT-PCR revealed decreased expression of NR1D1 in two primary BMAL1-KD CRC cell lines (HCT116 and SW480) but not in the metastatic CRC cell line SW620. (D) Quantitative RT-PCR revealed increased expression of CLOCK in two primary BMAL1-KD CRC cell lines but not in the metastatic CRC cell line SW620.