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. 2006 Jul 3;174(1):53–63. doi: 10.1083/jcb.200604058

Figure 1.

Figure 1.

Golgi biogenesis coincides with increased COPII assembly. (A–H) NRK cells were untreated (A and E) or treated with BFA/H89 (30 min BFA and 10 min H89) followed by the indicated times of washout to allow Golgi assembly. The methanol-fixed cells were costained with anti-giantin (A–D) and anti-Sec13 antibodies (E–H). Single optical sections are shown. Bar, 10 μm. (I) To quantify COPII assembly and ER exit of giantin over time, total above-threshold fluorescence levels of Sec13 and giantin per NRK cell were determined (mean ± SEM; >15 cells each). Coincident with giantin exit, COPII assembly increased before returning to steady-state levels for untreated cells (dashed lines). (J) Quantified COPII assembly based on GFP-Sec13 expression in HeLa cells is compared for a representative cell after BFA/H89 washout and two untreated control cells. Imaging was at 2-min intervals, and values are corrected and normalized to adjust for slight photobleaching and to allow direct comparison. (K) Means per cell for the number, area, and intensity of above-threshold Sec13 objects are compared over time during BFA + H89 washout in NRK cells. Note that increases in exit sites, i.e., object number, and mean size account for the total COPII assembly increase shown in I.