Fig. 2: Endogenous tagging and overexpression confirm assembly of TFG into anisotropic condensates that disassemble during mitosis.
From: A hollow TFG condensate spatially compartmentalizes the early secretory pathway
a Representative widefield micrograph of HeLa cells with homozygous endogenously tagged TFG::mClover-FLAG. Scale bar 10 μm. Magnified micrograph (dashed box in cell overview): individual TFG condensate. Scale bar 500 nm. b Quantification of the diameter of endogenous TFG condensates = 30. The dashed line represents the average of in vitro condensate diameter (341 nm ± 87) and the dotted line represents the average of endogenous ER/Golgi interface diameter (269 nm), for reference. c Qualitative fluorescence recovery after photobleaching (FRAP) of a TFG condensate. Scale bar 500 nm. d Left: live-cell confocal microscopy of large individual condensates within HeLa cells transfected with TFG-mEGFP-FLAG. Scale bar 5 μm. Middle: STED micrograph of large individual condensates within HeLa cells transfected with TFG-mEGFP-FLAG. Scale bar 2 μm. Right: 3D-structured illumination microscopy (3D-SIM) of large individual condensates within HeLa cells transfected with TFG-tGFP. Scale bar 500 nm. e Representative micrographs of partial and full FRAP experiments on overexpressed TFG condensates in HeLa cells. f Quantification of average condensate recovery rates for partial (n = 33) and full (n = 30) FRAP; normalized to pre-bleach (=100) and post-bleach intensity (=0). Error bars represent standard deviation. g Representative live-cell imaging of TFG condensate fusion dynamics. h Line scan of condensate depicted in (g). Source data are provided as a Source Data file.
