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. 2010 Mar 4;5(3):e9537. doi: 10.1371/journal.pone.0009537

Figure 3. The expanded Golgi remains intact.

Figure 3

A, HeLa cells transfected with control (ctrl) or Scyl1 miRNA and MannII-GFP were subjected to FRAP and FLIP analysis as described in the Materials and Methods. The images reveal MannII-GFP fluorescence with boxes indicating the photobleached areas. Images were collected prior to bleaching (pre), immediately after the first bleach (0) or at the indicated times (in seconds) after bleaching. Scale bars = 10 µm. B, The normalized intensity of fluorescence staining over the course of the FLIP and FRAP experiments. For the FLIP experiments (top graph) the y-axis value reflects the % GFP intensity normalized to the zero time fluorescence intensity prior to the repeated photobleachings during FLIP. Error bars are standard error of the mean (N = 9 for ctrl miRNA; N = 7 for unbleached Golgis and N = 6 for Scyl1 miRNA). No significant difference in the FLIP of Mannosidase II-GFP was observed between ctrl and Scyl1 miRNA expressing cells. For the FRAP experiments (bottom graph) the y-axis = the Golgi fluorescence intensity over the cytosolic fluorescence intensity during FRAP. There was no significance difference in the half maximal recovery time of Mannosidase II in either control of KD conditions. However, p115 KD eliminated Mannosidase II recovery by fragmenting the Golgi. Error bars represent standard error of the mean (ctrl miRNA N = 12; Scyl1 miRNA N = 9; p115 siRNA N = 8).