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. 2022 Jan 27;59(1):2100267. doi: 10.1183/13993003.00267-2021

FIGURE 2.

FIGURE 2

All surfactant protein C (SFTPC) is tracked from early compartments to the plasma membrane. a) Schematic of Retention Using Selective Hooks (RUSH) system, in which proteins are held in early tracking compartments using a streptavidin “hook” before release after addition of biotin. b) Localisation of SFTPC in HeLa cells expressing the RUSH-green fluorescent protein (GFP)-SFTPC fusion protein treated±biotin for 16 h confirms they traffic normally to post-Golgi compartments. c) Real-time tracking of GFP-SFTPC with transferrin receptor (TfnR) or CDMPR reveals co-localisation of SFTPC with TfnR in tubular structures, but failure of co-localisation beyond the Golgi with CDMPR which is seen in vesicles. d) After 2 h of biotin exposure, GFP-SFTPC-wild type (WT) is visible at the plasma membrane; this is reflected in increased BRICHOS domain presence at the plasma membrane by flow cytometry. SBP: streptavidin-binding protein, mCh: mCherry. Scale bars=10 μm.