Original article: DEXMEDETOMIDINE INHIBITS INFLAMMATION IN MICROGLIA CELLS UNDER STIMULATION OF LPS AND ATP BY C-FOS/NLRP3/CASPASE-1 CASCADES

Transcription

1 Supplementary material to Original article: DEXMEDETOMIDINE INHIBITS INFLAMMATION IN MICROGLIA CELLS UNDER STIMULATION OF LPS AND ATP BY C-FOS/NLRP3/CASPASE-1 CASCADES Hu Li 1, Xueping Zhang 2, Mingfu Chen 3, Jianyan Chen 4#, Tao Gao 3#, Shanglong Yao 5 1 Department of Anesthesiology, Shenzhen Baoan Hospital Affiliated to Southern Medical University, Shenzhen , China 2 Department of Anesthesiology, Shenzhen People s Hospital, The Second Clinical Medical College, Jinan University, Shenzhen Anesthesiology Engineering Center, Shenzhen , China 3 Department of Anesthesiology, Shenzhen Shajin Hospital Affiliated to Guangzhou Medical University, Shenzhen , China 4 Department of Anesthesiology, Second People's Hospital of Futian District, Shenzhen , China 5 Department of Anesthesiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan , China Both authors contributed equally to this work. # Corresponding authors, chenjianyan@yahoo.com (J. Chen), gaotao197771@163.com (T. Gao) This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( S1

2 Figure 1A: Cell viability tests Compared to control A B C C C *The tested value is absorbance. Test (1-3) indicates that one experiment was repeated for 3 times. Note: The absorbance in the table is equal to original test values deducting the average background value (0.023). For example in the group A of Test1, =0.466 Figure 1B: Extracellular concentrations of inflammatory factors IL-1β concentration Concentrations (pg/ml) A B C C C *The tested value is absorbance. Note: a, b, c, d, e, f, g and h indicate the test values of the standard substance of IL-1β at 500, 250, 125, 62.5, 31.25, , 7.8, and 0 pg/ml, respectively. The absorbance in the table is equal to original test values deducting the average background value (0.028). For example in the group A of Test1, =0.735 IL-18 concentration Concentrations (pg/ml) A B C C C *The tested value is absorbance. Note: a, b, c, d, e, f, g and h indicate the test values of the standard substance of IL-18 at 1000, 500, 250, 125, 62.5, 31.25, , and 0 pg/ml, respectively. The absorbance in the table is equal to original test values deducting the average background value (0.028). For example in the group A of Test1, =0.463 S2

3 Figure 1C: Relative protein levels NLRP3 protein level A B C C C *The value is the ratio of optical density of NLRP3 to that of GAPDH. Noncleaved caspase-1 A B C C C *The value is the ratio of optical density of noncleaved caspase-1 to that of GAPDH. Cleaved caspase-1 A B C C C *The value is the ratio of optical density of cleaved caspase-1 to that of GAPDH. S3

4 Figure 2A: Relative protein expression A A A A *The value is the ratio of optical density of c-fos to that of histone. Figure 2B: Relative luciferase activity A B C D E *The value is relative luciferase activity. S4

5 Figure 3A: c-fos protein levels in cell nucleus A B C D E *The value is the ratio of optical density of c-fos to that of histone. Figure 3A: Relative protein levels of NLRP3 A B C D E *The value is the ratio of optical density of NLRP3 to that of GAPDH. Figure 3B: Fluorescence intensity of c-fos A B C D E *The value is the fluorescence intensity. Figure 3C: Relative mrna levels of NLRP3 A B C D E *The value is relative mrna expression level of NLRP3. S5

6 S6

7 S7

8 S8

9 S9

10 S10

11 S11

12 S12