Functional Roles of Pattern Recognition Receptors That Recognize Virus Nucleic Acids in Human Adipose-Derived Mesenchymal Stem Cells

Figure 2

Poly(I:C)-induced immune responses. (a) Upregulation of TLR3 and RIG-I. hAD-MSCs were stimulated with 5 μg/mL poly(I:C) at indicated time. Relative mRNA levels of TLR3 and RIG-I were determined by real-time RCR at different time points. (b) The protein levels of TLR3 and RIG-I were determined by Western blot. hAD-MSCs were lysed 24 h after poly(I:C) stimulation. β-Actin was used as loading controls. (c) Poly(I:C) induced the expression of IFN-α and IFN-β in a time-dependent manner. Total RNA was extracted from hAD-MSCs, which were stimulated 5 μg/mL poly(I:C) in different time. Relative mRNA levels of IFN-α and IFN-β were determined using real-time PCR by normalizing to β-actin. (d) Poly(I:C) induced the expression of IFN-α and IFN-β in a dose-dependent manner. Total RNA were stimulated with the indicated dose of poly(I:C) for 6 h. Relative mRNA levels of IFN-α and IFN-β were determined by real-time PCR. (e) The secretion of IFN-α and IFN-β. hAD-MSCs were stimulated with poly(I:C). The expression levels of IFN-α and IFN-β in culture medium were measured by ELISA. (f) Expression of antiviral proteins in mRNA level. Total RNA was extracted from hAD-MSCs at the different time points after poly(I:C) stimulation. Relative mRNA levels of Mx1, OAS1, and ISG15 were determined using real-time PCR. (g) Expression of antiviral proteins in protein levels. The cell lysates of hAD-MSCs were to probe antiviral proteins by Western blot using specific antibodies after stimulation with poly(I:C) 24 h. The cells treated with LyoVec along served as control (Ctrl). Western blot images are representatives of at least three experiments. Data are presented as the mean ± SEM of three experiments. ; .

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