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. 2021 Oct 1;16(10):e0258111. doi: 10.1371/journal.pone.0258111

Fig 1. Protocol flow chart.

Fig 1

(A) Galactose-induced co-expression of the GPI-anchored cargo Gas1-GFP (green) and the transmembrane (TM) cargo Mid2-iRFP (blue) in the sec31-1 temperature-sensitive COPII mutant strain constitutively expressing the ERES marker Sec13-mCherry (red). (B) Accumulation of Gas1-GFP and Mid2-iRFP at the ER membrane by inhibiting COPII-coated vesicle formation at restrictive temperature (37°). (C) Restoration of ER export by shifting down to permissive temperature (24°C) and progressive incorporation of Gas1-GFP and Mid2-iRFP into the ERES. (D) Visualization of Gas1-GFP and Mid2-iRFP within the ERES by SCLIM microscopy.