Additional file1: of AMPK activation attenuates inflammatory pain through inhibiting NF-κB activation and IL-1β expression

Figure S1. Experimental design timeline. a. Nociceptive behavioral tests of AICAR effect in mice. Nociceptive behavioral tests were performed before (baseline) and 1 to 4 days after CFA injection. In the CFA plus AICAR group, rats received AICAR administration on 4th day after CFA injection. Ultimate effect of AICAR was measured at 0.5, 2, 4, 8, 12, 24, 48 h after the AICAR treatment. b. Biochemical experiments in mice. At day 4 of CFA injection, after 2 h of AICAR treatment, tissues for Western blotting and immunofluorescence labeling were collected in mice. c. Nociceptive behavioral tests of the effect of IL-1 ra in mice. Nociceptive behavioral tests were performed before (baseline) and 1 to 4 days after CFA injection. Behavior tests of ultimate effects of IL-1ra were performed at 0.5, 1, 2, 4, 8 h after IL-1ra treatment. d. Nociceptive behavioral tests of effects of Compound C on AICAR in CFA injection mice. e. Biochemical experiments in wild-type mice and CX3CR1-GFP mice. At day 4 of CFA injection, after 2 h of AICAR treatment (Compound C administration in the 30 min of AICAR), tissues for Western blotting were collected in wild-type mice, and tissues for immunofluorescence labeling were collected in CX3CR1-GFP mice. f. To determine whether knockdown of AMPKα may reverse AICAR effects, mice were injected with AMPKα shRNA Lentiviral Particles at CFA injection day 1. Behavior tests of ultimate effect of AICAR were performed at 1, 2, 4, 8 h after AICAR treatment at day 4. Tissues for Western blotting were collected after nociceptive behavior tests. Abbreviations: WB, Western blotting; IF, Immunofluorescence labeling. (PDF 8021 kb