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. 2020 Aug 5;12(8):2188. doi: 10.3390/cancers12082188

Figure 2.

Figure 2

YB-1 was required for TNF-induced NF-κB activation. (A) Bone marrow-derived macrophages (BMDM, WT, and YB-1 KO) were stimulated with TNF (20 ng/mL) for the indication of time periods to activate the NF-κB signaling pathway. Protein expression was analyzed by immunoblotting using the indicated antibodies. Vinculin was used as the loading control. The relative band intensities are indicated. (B) Relative YB-1 expression is shown for three independent experiments. WT (white) and KO (black bars). (C) Control and YB-1 knockdown (KD) cells (THP-1 and U937) were stimulated with TNF (20 ng/mL) to activate tumor necrosis factor receptor 1 (TNFR1). Proteins were analyzed by immunoblotting for expression of the indicated proteins. Vinculin was used as loading control. Control (CTRL; white) and knockdown (KD; black bars). (D) Relative YB-1 expression in KD cells is shown for three independent experiments. (E) Relative pp65, phospho- inhibitor of NF-κB alpha (p-IκBα), phospho-IkB kinase α/β (pIKKα/β), and TNF receptor-associated factor 2 (TRAF2) expression in WT and KD THP-1 and U937 cells are shown for three independent experiments. Control (CTRL; white) and knockdown (KD; black bars). Error bars specify the standard error of the mean (SEM). Statistical significance was calculated using an unpaired t-test, n = 3. Data represent the mean ± SEM. * p < 0.05, ** p < 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.