Fig. 8
From: Cordycepin prevents radiation ulcer by inhibiting cell senescence via NRF2 and AMPK in rodents
Cordycepin interacts with α1 and γ1 subunit of AMPK. a Akt, p-Akt, S6, p-S6 expression levels from 200 μM cordycepin-treated fibroblasts for the indicated times. b Representative immunofluorescence pictures of p-S6 of the tongues from normal mice and irradiated mice (control, prevention, treatment) 10 days after irradiation dose. c AMPK, p-AMPK, ACC, p-ACC expression levels from 200 μM cordycepin-treated fibroblasts for the indicated times. This test was repeated three times. d Fibroblasts were incubated in the absence or presence of 40 μM Compound C (C.C) for 4 h in advance and then were treated with 200 μM cordycepin for the indicated times (kept with C.C), and total proteins were harvested for detection of p-AMPK, S6, p-S6, NRF2, KEAP1, and LC3 by western blot. e Analysis of reactive oxygen species (ROS) levels 24 h after radiation in fibroblasts, fibroblasts were incubated in the absence or presence of 40 μM C.C for 4 h in advance and then were treated with 200 μM cordycepin for 3 days (kept with C.C) before radiation. f Representative images of fibroblasts colonies generated in survival assays, fibroblasts were incubated in the absence or presence of 40 μM C.C for 4 h in advance and then were treated with 200 μM cordycepin for 3 days (kept with C.C) before radiation. g p16 and p21 expression levels from fibroblast 7 days after radiation, fibroblasts were incubated in the absence or presence of 40 μM C.C for 4 h in advance and then were treated with 200 μM cordycepin for 3 days (kept with C.C) before irradiation. h Effect of cordycepin on intracellular AMP, ADP, ATP levels, and the AMP/ATP ratio, fibroblasts were treated with 200 μM cordycepin for the indicated times. i Molecular docking study of the binding affinity of cordycepin to AMPK. j Fibroblasts were transfected with control siRNA (si-Ctl), siRNA against AMPK-α1 (si-α1) or siRNA against AMPK-γ1 (si-γ1), and then were treated with 200 μM cordycepin for 3 days, and total proteins were harvested for detection of S6, p-S6 and NRF2 by western blot. Bars represent 50 μm (b). Data in e and h represent the means ± S.D. (n = 3, **P < 0.01; student’s t-test; ns not significant)
