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. 2019 Sep 11;8(9):1064. doi: 10.3390/cells8091064

Figure 3.

Figure 3

The characteristic of RAW264.7 cell after LPS stimulation once (two doses; 100 and 200 ng/well) or twice, LPS/LPS (100/100), versus control (N/N) (see method) as determined by supernatant cytokines (A–C) and Lipocalin-2 (Lcn-2; D) were demonstrated. Additionally, Lcn-2 receptor (24p3R) after cell stimulations by Western blot analysis (E) and NFκB, a possible downstream signaling of Lcn-2, in cells with or without recombinant Lcn-2 (rLcn-2) activation by relative mRNA expression (F) were indicated. Moreover, cell viability as determined by nuclear staining as calculated by ImageJ (G), representative nuclear stain figures (H–J) and metabolic activity (MTT assay, see method) (K) (see method) were determined. Further, extracellular flux analysis of RAW264.7 cell with LPS/LPS versus control as demonstrated by oxygen consumption rate of mitochondria stress test (L) and extra-cellular acidification rate of glucose stress test (M) were demonstrated. (Independent triplicate experiments were performed for all experiments; *, p < 0.05; **, p < 0.01; #, p < 0.01 vs. N/N); Max. Resp. Capacity, maximum respiratory capacity area.