Expression of functional IL-27 heterodimeric receptor on FLS. (A, B). Total RNA was extracted from control FLS, RA-FLS and PBMC (positive control), followed by quantitiatve real time RT-PCR analysis for gp130, WSX-1 and GAPDH (house keeping gene) expression. (C) Representative histograms of cell surface expression of gp130 on control FLS, RA-FLS and human CD4+T cells (positive control) determined by flow cytometry. (D) Representative Western blot analysis of WSX-1 protein expression of control FLS, RA-FLS and PBMC (positive control). GAPDH was used as protein control to ensure an equal amount of loaded protein. All the experiments were performed in three independent replicates.
M, 100 base-pair molecular size marker; PBMC, peripheral blood mononuclear cells.