Fig 2: Intracellular ROS generation upon lipolysis regulation. (a) Measurement of intracellular ROS. Cells were treated with 5 µM of SU6656 (SU) in the presence or absence of 20 µM of FABP4 inhibitor HTS01037 (HTS) for 3 days and followed by incubation with 1 µM of CM-H2DCF-DA for H1993 or 2·5 µM of CM-H2DCF-DA for Calu6 cells. Pictures were taken using confocal microscope followed by quantitative analysis using MetaMorph 6.3 software (Molecular Devices). Scale bar = 20 µm (b) AMPK phosphorylation upon inhibitor treatment. Cells treated with SU or HTS for 3 days were assayed for AMPK and pAMPK expression in H1993 and Calu6 cells. (c) Oxygen consumption rates in lung cancer cell lines with inhibitor treatment. Calu6 and H1993 cells were treated with SU, HTS or both for 3 days. The OCRs were measured as in materials and methods. (d) Cell growth assay with inhibitor treatment. Cell proliferation and MTT assays show cell viability under SU, HTS, or both treatment for 3 days. (e) Protein expression involved in regulation of cell cycle or apoptosis in Calu6 and H1993 upon indicated treatment for 24 h. Values are mean ± SEM. Results represent data from at least two independent experiments. Asterisks refer to *P = 0·05; **P = 0·01; ****P = 0·0001 (One-way ANOVA, Tukey's post test).

Fig 3: Effect of PDX on adenosine monophosphate-activated protein kinase (AMPK) signal pathway in spinal cord horns (n = 3/group). Western blot (A) and immunohistochemistry (C) were respectively conducted to evaluate the alterations in the AMPK signaling pathway. Relative to sham group, the protein levels of expression of p-AMPK markedly were suppressed in model rats. Meanwhile, the treatment of PDX could dose-dependently reverse the upper alterations of p-AMPK. Statistical analysis of p-AMPK (B,D). One-way ANOVA was used for comparisons among groups followed by Turkey’s multiple comparisons test; Values are displayed as mean ± SEM (*p < 0.05 vs. sham group; #p < 0.05 vs. vehicle group; ^p < 0.05 vs. 10 ng group).

Fig 4: Activation of autophagy. AMPK is similar between C57 and mdx mice at both 7 wk and 17 mo of age. Significance was established at p < 0.05. White bars – C57 7 wk (n = 4–5), gray bars – mdx 7 wk (n = 7–8), white bars with black hatching – C57 17 mo (n = 3), black bars – mdx 17 mo (n = 8).

Fig 5: Schematic diagram showing the effect of aucubin on autophagy activation in dexamethasone-induced osteoblasts: The autophagy of osteoblasts is maintained at basal levels under normal conditions and is increased when it faces excessive steroids to achieve self-protection. However, insufficient autophagy activation fails to prevent osteoblasts apoptosis, eventually leading to osteonecrosis. Aucubin enhances the level of autophagy and effectively prevents steroid-induced osteoblasts apoptosis by activating AMPK signalling