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. Author manuscript; available in PMC: 2011 Sep 3.
Published in final edited form as: Biochem Biophys Res Commun. 2010 Jul 30;399(4):498–504. doi: 10.1016/j.bbrc.2010.07.096

Fig. 3. The Golgi-to-ER retrograde trafficking regulates EGF-dependent EGFR transport to the nucleus.

Fig. 3

(A) Pretreatment with BFA inhibited EGF-dependent EGFR translocation to the nucleus in MDA-MB-468 cells. Cells maintained in a serum-starved medium for 24 hr were pretreated with BFA (5 μg/ml) for 30 min and then treated with or without EGF, followed by cellular fractionation and subjected to immunoblotting with the indicated antibodies. (B and C) The dominant ARF1mutants inhibited EGF-dependent EGFR translocation to the nucleus. Cells were transfected with ARF1/wt or two ARF1 mutants (ARF1/T31N and ARF1/Q71L) and then treated with or without EGF. In (B), proteins from the non-nuclear fractions (non-N, odd-numbered lanes) and nuclear fractions (N, even-numbered lanes) by cellular fractionation were immunoblotted with the indicated antibodies. In (C), cells were immunostained with EGFR and analyzed using confocal microscopy. Bar, 5 μm. The bar diagram indicates detection of nuclear localization of EGFR calculated for a total of 50 cells.