Murine Pancreatic Beta TC3 Cells Show Greater 2^′,5^′-Oligoadenylate Synthetase (2^′5^′AS) Antiviral Enzyme Activity and Apoptosis Following IFN-𝛼 or Poly(I:C) Treatment than Pancreatic Alpha TC3 Cells

Figure 1

Protein expression and activity of 2^′5^′AS in non-stimulated and IFN-α stimulated beta TC3 and alpha TC3 cells. After 24 hours of IFN-α stimulation, cell lysates were prepared. (a) For immunoblot, cell extracts were separated on 12% SDS-PAGE and transferred to nitrocellulose membranes. The polyclonal antibody specific to human 2^′5^′AS (cross-reactive with murine 2^′5^′AS) was used to detect the expressed proteins. The polyclonal antibody against mouse GAPDH was used to quantitate the loading amounts; (b) For assay of 2^′5^′AS activity, lysates were incubated with 0.15 μCi α ³²P-ATP for 120 minutes at 30^∘C in the presence of 100 𝜇g/ml poly(I:C), and then separated by ascending TLC. The generated 2^′5^′-oligoadenylates (2^′5^′A) were quantitated by PhosphorImager analysis. Enzyme activity was expressed in units per milligram protein (𝑛=3).

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