During cER to PM membrane transport, Ist2 does not get delivered to the cell surface. Wild-type cells expressing GFP-Ist2 and the ER marker ER-RFP (SEY6210 pMT1 pMT10) were spheroplasted, flushed into the microfluidics chamber, and exposed to hypoosmotic shock (0.4 M sorbitol > 0.1 M sorbitol). (A and B) Subpart (A) shows fluorescence microscopy pictures of the surface and cross section of one of these spheroplasts and (B) shows the surface of two additional examples. The star marks cell debris (not part of the analyzed cell).