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. 2018 Jun 1;9(42):26884–26899. doi: 10.18632/oncotarget.25529

Figure 1. Analysis of the expression of TNFAIP8 isoforms in cancer cell lines.

Figure 1

(A) Amino acid sequence alignment of the five protein isoforms of human TNFAIP8. The N-terminal region is variable, whereas the C-terminal region is highly conserved. (B) Total RNA from PC3, MDA-MB-468, LNCaP, and LLC1 cells were isolated and the expression of the five TNFAIP8 isoforms was analyzed using RT-PCR. PCR products were electrophoresed on a 1.5% agarose gel and stained with ethidium bromide. NC, negative control (no cDNA); M, markers. (C) Expression of TNFAIP8 protein isoforms in various types of cancer cell lines was analyzed by immunoblotting. Cell extracts (50 μg) were electrophoresed using SDS-PAGE and immunoblotted with anti-TNFAIP8 and anti-β-actin antibodies. (D) PC3, LNCaP, and C4-2 cells were cultured for 24 h and treated with TNFα (10 ng/ml) for 3 h and 15 h. Cell extracts (50 μg) were immunoblotted with anti-TNFAIP8 and anti-GAPDH antibodies. (E) PC3 cells were transfected with control siRNA or TNFAIP8 siRNA (100 nM) for 30 h. Cell extracts (50 μg) were immunoblotted with anti-TNFAIP8 and anti-GAPDH antibodies. Endo–endogenous.