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. 2021 Sep 22;16(9):e0257576. doi: 10.1371/journal.pone.0257576

Fig 2. 25HC is required for optimal TNF mediated pro-inflammatory response and NFκB activation.

Fig 2

(A) Medium supernatant collected from WT and C25H KO BMDMs treated with TNF (100ng/ml) for 16h was analyzed for CCL3 production by ELISA (n = 12; three independent experiments). (B) Cell lysate collected from WT and C25H KO cells treated with TNF (100ng/ml) for 0–60 minutes were subjected to western blot analysis to detect status of phosphorylated IκB (phospho-IκB). The immunoblot is representative of three independent experiments with similar results. (C) Densitometric analysis of the western blot presented in Fig 2b. The densitometric quantification values for phospho-IκB immunoblot represent the ratio of phospho-IκB:actin and the fold-induction was calculated after normalizing with the control 0 min group. ELISA data are shown as Mean ± SEM. *p ≤ 0.05 using a Student’s t-test. The densitometric values represent the mean ± SEM from three independent experiments. The p value shown in the figure was calculated using a Student’s t-test (*p ≤ 0.05).