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Figure 1. Validation of BCP rat models (A) Radiological images of the MRMT-1-injected and Hank’s buffer-injected left hindpaw of the rats, 21 days after.

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Presentation on theme: "Figure 1. Validation of BCP rat models (A) Radiological images of the MRMT-1-injected and Hank’s buffer-injected left hindpaw of the rats, 21 days after."— Presentation transcript:

1 Figure 1. Validation of BCP rat models (A) Radiological images of the MRMT-1-injected and Hank’s buffer-injected left hindpaw of the rats, 21 days after inoculation. No effect was seen on the buffer-injected left hindpaw. Arrows indicate the site of injection. (B) Mechanical PWT was tested using a series of calibrated von Frey filaments prior to tumor cell inoculation (Day 0) and on POD 4, 7, 10, 12, 14, 17, 21, and 24. ■: Sham group, ○: model group. All data were expressed as the mean ± SEM, n = 9. *P < 0.05, **P < 0.01 vs the Hank’s group rats at each corresponding time point. (C) Western blot analysis of ASIC3 protein level in the ipsilateral L4–L6 DRG of sham and BCP model rats on POD 21. (D) Image J software were used to quantify the gray degree values. The results were shown as the mean ± SD. Three independent experiments were performed. From: Resveratrol attenuates bone cancer pain through regulating the expression levels of ASIC3 and activating cell autophagy Acta Biochim Biophys Sin (Shanghai). Published online October 03, doi: /abbs/gmx103 Acta Biochim Biophys Sin (Shanghai) | © The Author Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please

2 Figure 2. Effect of resveratrol on the BCP and the acidosis-evoked pain (A) PWT of BCP rats on POD 21 after intraperitoneal injection of vehicle (DMSO + saline), 0.3, 1 and 3 mg/kg resveratrol. (□: vehicle group, red ●: 0.3 mg/kg resveratrol group, green ▲: 1 mg/kg resveratrol group, and blue ▼: 3 mg/kg resveratrol group). All data were expressed as the mean ± SEM, n = 9. (*P < 0.05, **P < 0.01 vs the vehicle group rats at each corresponding time point). (B) PWT of normal rats after intraperitoneal injection of vehicle (DMSO + saline) and 3 mg/kg resveratrol. (□: vehicle group, red ●: 3 mg/kg resveratrol group). All data were expressed as the mean ± SEM, n = 9. (C) The flinch/shaking response of rats in the acidosis-evoked pain after intraperitoneal injection of vehicle (DMSO + saline) and 3 mg/kg resveratrol. Flinching shaking of paw was recorded as the number of flinches per observation period (5 min). **P < 0.01, unpaired t-test, compared with control column. n = 9. From: Resveratrol attenuates bone cancer pain through regulating the expression levels of ASIC3 and activating cell autophagy Acta Biochim Biophys Sin (Shanghai). Published online October 03, doi: /abbs/gmx103 Acta Biochim Biophys Sin (Shanghai) | © The Author Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please

3 Figure 3. The effect of resveratrol on the expression level of ASIC3 protein (A) Western blot analysis of GFP-ASIC3 protein level in total lysate of SH-SY5Y cells exposed to resveratrol (0, 0.1, and 1 μM) for 24 h. (B) Western blot analysis of GFP-ASIC3 protein level in the membrane of SH-SY5Y cells exposed to resveratrol (0, 0.1, and 1 μM) for 24 h. (C) Western blot analysis of GFP-ASIC3 protein level in total lysate of SH-SY5Y cells exposed to 1 μM resveratrol for 0, 12, and 24 h. (D–F) Image J software were used to quantify the gray degree values. The results were shown as the mean ± SD. Three independent experiments were performed, *P < 0.05, **P < 0.01. From: Resveratrol attenuates bone cancer pain through regulating the expression levels of ASIC3 and activating cell autophagy Acta Biochim Biophys Sin (Shanghai). Published online October 03, doi: /abbs/gmx103 Acta Biochim Biophys Sin (Shanghai) | © The Author Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please

4 Figure 4. The effect of resveratrol on the cell autophagy signal pathway (A) Western blot analysis of autophagy associated protein expression levels after exposure to 1 μM resveratrol for 24 h in ASIC3-transfected SH-SY5Y cells. (B,C) Flow cytometric analysis of Annexin V and PI staining for ASIC3-transfected SH-SY5Y cells treated with DMSO and 1 μM resveratrol for 24 h, respectively. The percentage of apoptotic cells was detected by analyzing Annexin V-FITC and PI binding with the help of the Submit 4.0 software. Viable cells do not bind with Annexin V or PI (lower left quadrant D3), early apoptotic cells bind with Annexin V but excludes PI (lower right quadrant D4), necrotic or late apoptotic cells are both Annexin V and PI-positive (upper right quadrant D2). The upper left quadrant D1 contains cells damaged during the preparation of the cell suspension. (D) Western blot analysis of SIRT1 and LC3 protein expression levels in the L4–L6 DRG of sham and BCP rats on POD 21. (E) Western blot analysis of ASIC3 protein expression level transfected with or without SIRT1 for 24 h in SH-SY5Y cells. From: Resveratrol attenuates bone cancer pain through regulating the expression levels of ASIC3 and activating cell autophagy Acta Biochim Biophys Sin (Shanghai). Published online October 03, doi: /abbs/gmx103 Acta Biochim Biophys Sin (Shanghai) | © The Author Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please

5 Figure 5. Hypothetical working model for the antinociceptive effect of resveratrol in BCP rats
From: Resveratrol attenuates bone cancer pain through regulating the expression levels of ASIC3 and activating cell autophagy Acta Biochim Biophys Sin (Shanghai). Published online October 03, doi: /abbs/gmx103 Acta Biochim Biophys Sin (Shanghai) | © The Author Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please

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