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. 2022 Dec 15;23(24):15959. doi: 10.3390/ijms232415959

Figure 8.

Figure 8

CEN-induced autophagy in SAOS400 cells was related to intracellular ATP levels and AMPK activation. Panel (a): SAOS400 cells were treated for 24 h with vehicle (Veh), CEN, 5 Gy, or a combination of 5 Gy plus CEN. Subsequently, the intracellular concentration (nmol) of ATP was measured with a luminometer as described in the Materials and Methods Section. The bar graphs indicate the mean of three experiments (±SD) in quadruplicate. Symbols: + p < 0.05 Veh vs. CEN, *** p < 0.001 5 Gy and CEN vs. 5 Gy, Student’s t test. Panel (b): Immunoblot analysis for the expression of the active form of AMPK (pAMPKαThr172). Cells were irradiated with 5 Gy and then treated with 200 μg/mL of CEN or their combination for 24 h. Values between panels (mean of two experiments with statistical significance: + p < 0.05 Veh vs. CEN, * p < 0.05 CEN vs. 5 Gy, Student’s t test) indicate a densitometric analysis of the expression, as described in the Materials and Methods Section. Panel (c): SAOS400 cells were irradiated (5 Gy) or incubated with the AMPK activator, AICAR (5 μM), or their combination. Cell viability was evaluated using Cy-Quant dye after 120 h of treatment. The bar graphs represent the mean ± SD, and symbols and the linear bars indicate significance: ++ p < 0.01 Ctrl vs. 5 Gy or AICAR, *** p < 0.001 5 Gy and AICAR single treatments vs. combined treatment AICAR + 5 Gy (Student’s t test).