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. Author manuscript; available in PMC: 2012 Feb 15.
Published in final edited form as: J Immunol. 2011 Jan 17;186(4):2164–2171. doi: 10.4049/jimmunol.1003236

Fig. 3. PTB is actively involved in the distribution of CD40L mRNA between the nucleus and cytoplasm.

Fig. 3

(A). Levels of CD40L mRNA levels in the cytoplasm (grey bars) and nucleus (black bars) of pLV-CTRL and pLV-PTB infected Jurkat/D1.1 and Flag-PTB-Jurkat/D1.1 cells as determined by quantitative PCR. Results represent the Average and SEM of 3 independent experiments. (B) Quantitative PCR analysis of CD40L mRNA from the non-polysomal (white bars) and polysomal (black bars) fractions of pLV-CTRL- and pLV-PTB-infected Jurkat/D1.1 cells and Flag-PTB-Jurkat/D1.1. (C). Western blot analysis using cytoplasmic (2 × 103 cell equivalents), nuclear (2 × 103 cell equivalents) and polysomal (1 × 104 cell equivalents) extracts of Jurkat/D1.1 (lanes 1-6) and Flag-PTB-Jurkat/D1.1 (lanes 7-12) cells infected with pVL-PTB or pVL-CTRL were analyzed for expression of PTB and nucleolin in the different cellular fractions by Western blotting. Membranes were re-probed with antibodies against β-actin and the cytoplasmic ribosomal protein S6 to validate the efficient separation of nuclear, cytoplasmic and polysomal fractions. The membrane was further hybridized with anti-Flag antibodies to confirm expression of the Flag-PTB in the different fractions. Boxed lanes show the effect of PTB downregulation on the different proteins, in particular PTB and nucleolin.