CK2 upregulated and induced SIRT1 and AMPK activation during calorie restriction in human cancer cells. (A) HCT116 and MCF-7 cells were incubated under CR conditions for the indicated time. (B and C) Cells were transfected with CK2α siRNA for 1.5 days, then incubated under CR conditions for 7 h. The levels of each protein (A and B) and mRNA (C) were then determined by immunoblotting using specific antibodies, and by RT-PCR analysis using specific primers, respectively (top). Representative data from three independent experiments are shown. β-Actin was used as a control. The graphs represent the quantitation of CK2α, CK2β, and SIRT1 relative to β-actin, and p-AMPK (T172) relative to the unphosphorylated AMPK (bottom). Data were reported as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. Bars that do not have the same letter (a, b, c) were significantly different among groups at P < 0.05.|@|~(^,^)~|@|CK2 positively regulated autophagy in human cancer cells. HCT116 and MCF-7 cells were transfected with CK2α siRNA or pcDNA3.1-HA-CK2α for 2 days. (A) Immunoblotting was then used to determine the level of each protein using specific antibodies (top). (B) The level of each mRNA was determined by RT-PCR using specific primers (top). Representative data from three independent experiments are shown. β-Actin was used as a control. The graphs represent quantitation of each protein and mRNA relative to β-actin (bottom). Data were reported as mean ± SEM. Bars that do not have the same letter (a, b, c) were significantly different among groups at P < 0.05.|@|~(^,^)~|@|CK2 positively regulated autophagy via mTOR and SIRT1 in human cancer cells. (A) HCT116 and MCF-7 cells were transfected with CK2α siRNA for 2 days in the absence or presence of 10 μM rapamycin or 10 μM resveratrol. The levels of each protein (left-top) and mRNA (left-bottom) were determined by immunoblotting using specific antibodies and by RT-PCR using specific primers, respectively. Representative data from three independent experiments are shown. β-Actin was used as a control. The graphs represent the quantitation of each protein (right-top) and mRNA (right-bottom) relative to β-actin. WB, Western blotting. (B) Cells were transfected with CK2α siRNA for 2 days in the absence or presence of pECE-flag-SIRT1 or pECE-flag-SIRT1 mutant (H363Y). The levels of each protein (left-top) and mRNA (left-bottom) were determined by immunoblotting using specific antibodies and by RT-PCR using specific primers, respectively. Representative data from three independent experiments are shown. β-Actin was used as a control. The graphs represent the quantitation of each protein (right-top) and mRNA (right-bottom) relative to β-actin. Data are reported as mean ± SEM. Bars that do not have the same letter (a, b, c) were significantly different among groups at P < 0.05.|@|~(^,^)~|@|CK2 positively regulated autophagy via AMPK and AKT pathways in human cancer cells. HCT116 and MCF-7 cells were transfected with CK2α siRNA for 2 days in the absence or presence of 10 μM AICAR or 10 μM triciribine. (A) The level of each protein was determined by immunoblotting using specific antibodies (top). (B) The level of each mRNA was determined by RT-PCR using specific primers (top). Representative data from three independent experiments are shown. β-Actin was used as a control. The graphs represent the quantitation of each protein and mRNA relative to β-actin (bottom). Data are reported as mean ± SEM. Bars that do not have the same letter (a, b, c) were significantly different among groups at P < 0.05.|@|~(^,^)~|@|Overexpression of FoxO3a rescued CK2 downregulation-mediated autophagy inhibition in human cancer cells. HCT116 and MCF-7 cells were transfected with CK2α siRNA for 2 days in the absence or presence of pECE-HA-FoxO3a. (A) The level of each protein was determined by immunoblotting using specific antibodies (top). (B) The level of each mRNA was determined by RT-PCR using specific primers (top). Representative data from three independent experiments are shown. β-Actin was used as a control. The graphs represent the quantitation of each protein and mRNA relative to β-actin (bottom). Data are reported as mean ± SEM. Bars that do not have the same letter (a, b, c) were significantly different among groups at P < 0.05.|@|~(^,^)~|@|Kin-10 RNAi decreased autophagy markers in nematodes. (A) Synchronized (L4 larva) reporter nematodes lgg-1::gfp were fed on control RNAi or kin-10 RNAi plates containing AICAR (100 μM), triciribin (TCN, 0.2 μM), or resveratrol (RSV, 50 μM) under standard conditions. Representative fluorescence images at day 1 of adulthood were obtained at 10× magnification (top). The fluorescence intensity of GFP was quantified using ImageJ software by determining the average pixel intensity (n = 30 per condition). (B) Synchronized (L4 larva) nematodes were fed on control RNAi or kin-10 RNAi plates containing AICAR (100 μM), triciribine (TCN, 0.2 μM), resveratrol (RSV, 50 μM), or spermine (SPM, 0.2 μM) under standard conditions. Subsequently, lysates from nematodes were used in RT-PCR using specific primers. The level of each mRNA was determined by RT-PCR (top). Representative data from the three independent experiments are shown. Act-2 was used as a control. The graphs represent the quantitation of each protein and mRNA relative to act-2 (bottom). Data are reported as mean ± SEM. Bars that do not have the same letter (a, b, c) were significantly different among groups at P < 0.05.|@|~(^,^)~|@|Schematic of the mechanism proposed for the calorie restriction-mediated autophagy induction.

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