Fig. 1
Authentic TAMPs integrate into membrane. a Representative confocal microscope images of NMuMG cells expressing the EGFP-tagged putative TA proteins (NP_057648, NP_064564, and NP_149991) and mLumin-tagged NP_079504 and NP_003797. Cells are co-stained with Mitotracker Red (EGFP-NP_057648, EGFP-NP_064564, and EGFP-NP_149991) and Mitotracker Green (mLumin-NP_079504 and mLumin-NP_003797) and nuclear counterstain DRAQ5. Scale bar = 25 μm. b By sodium carbonate extraction selected putative TA proteins were tested for membrane integration. Total (T) cell lysate prepared from NMuMG cells stably expressing the indicated EGFP fusion proteins was fractionated into a supernatant fraction (S) containing cytosolic proteins and fractions containing peripheral proteins (CS) and sodium carbonate resistant integral membrane proteins (CP). Aliquots were analyzed by immunoblotting using antibodies to GFP or RFP (mLumin)