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. Author manuscript; available in PMC: 2020 Jun 25.
Published in final edited form as: Eur J Cell Biol. 2010 Apr 1;89(7):564–573. doi: 10.1016/j.ejcb.2009.11.029

Fig. 6. B-ZIP|A-ZIP interactions in vivo by FRAP and FRET.

Fig. 6.

A) NIH3T3 cells were transiently co-transfected with GFP-VBP and 5 different mCherry-A-ZIPs and analyzed by FRAP. mCh-A-C/EBPα and mCh-A-C/EBPβ do not affect the recovery of GFP-VBP. mCh-A-VBP, mCh-A-CREB and mCh-A-cJun increase the recovery of GFP-VBP. B) NIH3T3 cells were transiently co-transfected with GFP-VBP-LZ and different mCh-LZ domains and analyzed by FRET. Cells were prepared for live cell imaging as in FRAP, and a Zeiss 510 confocal microscope with the same objective, incubation and GFP excitation conditions defined for FRAP was also used for FRET. mCherry excitation was achieved with the 543 laser line of a 1 mW HeNe laser, and mCherry emission was monitored with an LP560 filter above 560 nm. Cells were imaged in multi-track mode at 0.75% argon laser power and 25% HeNe laser power. Sensitized emission was used to detect FRET. The FRET efficiency between GFP-VBP-LZ with mCh-VBP-LZ homodimers was significantly greater than all other leucine zipper heterodimers (*p<0.05, **p<0.01). FRAP and FRET results are representative of at least 3 independent experiments in which at least 10–15 different cells were imaged in each experimental condition.