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Fig. 6 | Journal of Inflammation

Fig. 6

From: PKM2/STAT1-mediated PD-L1 upregulation on neutrophils during sepsis promotes neutrophil organ accumulation by serving an anti-apoptotic role

Fig. 6

STAT1 activation promotes PD-L1 upregulation and inhibits dHL-60 apoptosis under LPS stimulation. A-D dHL-60s were treated with LPS (1ug/ml) alone for 6 h, pretreated with Fludarabine (50 and 100 uM) before LPS stimulation or treated with Fludarabine (100uM) alone. A, B Protein level of p-STAT1 was determined by Western blotting. β-actin was used as the loading control. The blot is representative of three independent experiments. C, D PD-L1 levels on dHL-60s under different treatments were measured by flow cytometry. Proportion of PD-L1+ dHL-60 s (C) and MFI of PD-L1 on dHL-60s (D) were measured (n = 3). E-I dHL-60 s were treated with LPS (1ug/ml) alone, pretreated with Fludarabine (50uM) before LPS stimulation or treated with Fludarabine (50uM) alone. E, F dHL-60s were stained by annexin V/PI and assessed by flowcytometry (n = 3). E Representative flow cytometry images of dHL-60 apoptosis. F Histogram of the apoptosis rates of dHL-60 s subjected to different treatments. G-I Levels of apoptosis-related proteins were determined by Western blotting. Hsp90 was used as the loading control. The blot is representative of three independent experiments. One-way ANOVA was performed. Data are presented as the mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001

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