Figure 1: Detections of A2AR effect on apoptosis and autophagy of bone marrow cells and peripheral neutrophils in LPS-induced SIRS model. | Scientific Reports

Figure 1: Detections of A2AR effect on apoptosis and autophagy of bone marrow cells and peripheral neutrophils in LPS-induced SIRS model.

From: Activation of Adenosine 2A receptor inhibits neutrophil apoptosis in an autophagy-dependent manner in mice with systemic inflammatory response syndrome

Figure 1

(A–C) Evaluations of SIRS related indexes including rectal temperature (A), breath frequency (B) after LPS intraperitoneal injection (11.25 million EU/kg, 10 mg/Kg) for 6 hours. The SIRS severity scores (C) were counted after LPS injection for 6 hours and 24 hours. Eaqual saline was injected into WT/A2AR KO mice as control. White bars represent before injection and gilled bars represent after injection for 6 hours, black bars represent severtiy score after 24 hours development. (D) Survival rates of each group. Each group contained 5 mice treated with intended conditions. The mortality of each group was counted until 24 hours’ development. (E) Flow cytometry analyse of apoptosis cells on bone marrow cells which were harvested after LPS injection for 24 hours. The harvested bone marrow cells were stained with AnnexinV/7-AAD and analyzed using MUSE cell analyzer. (F) The quantification of total apoptotic bone marrow cells in (E). (G) Detection of apoptosis neutrophils isolated from mice peripheral blood. The sample slides were stained by Apoptag flurorescein to detect the apoptosis cells, as the white arrows show. The nucleus were stained by DAPI. The upper white line represents 100 μm and the lower white line represents 50 μm. (H) The counted proportions of total apoptotic cells in mice peripheral neutrophils in (G). (I) Assays of autophagy related molecule LC3 by WB. The protein lysis were achieved from isolated bone marrow cells of above four groups. The ratio of LC3/GAPDH was calculated to reflect the level of autophagy. (J) The arbitrary units were shown as the densitometry ratio of LC3 compared with GAPDH. Each mice group (except the survival experiment in D contained 5 animals) contained 3 animals which were with similar weight and age. Data represents the mean ± SEM for three independent experiments. (*p < 0.05 between the two groups; **p < 0.01 between the two groups, NS, no significant difference between the two groups, Student’s t test).

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