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Fig. 2 | BMC Plant Biology

Fig. 2

From: NtGNL1a ARF-GEF acts in endocytosis in tobacco cells

Fig. 2

Endomembrane trafficking inhibitors, auxins and cytoskeletal drugs interfere with formation of BFA-induced PIN1-GFP aggregations. a-l In vivo confocal microscopy of 3-day-old tobacco BY-2 cells transformed with PIN1::PIN1:GFP after 30 min pre-treatment with individual inhibitors, auxins and cytoskeletal drugs followed by 30 min with 20 μM BFA added. Confocal sections of perinuclear plane captured with confocal microscope (488 nm excitation). a CTRL cells with mock treatment (DMSO), note PIN1-GFP at the PM. b 30 min of BFA (20 μM) treatment inducing intracellular aggregations of PIN1-GFP. c TYR A23 (50 μM) preventing the BFA-induced PIN1-GFP intracellular aggregations in contrast to (d) TYR A51 (50 μM) pre-treatment. e WM (33 μM), note that BFA-induced PIN1-GFP aggregations differ from the BFA effect alone. f FIL (15 μM) and (g) DNS (80 μM) partially inhibiting the BFA-induced PIN1-GFP intracellular aggregations, note less PIN1-GFP in the cytoplasm. j 2,4-D (5 μM) inhibition of BFA-induced PIN1-GFP aggregations (i) in contrast to IAA (5 μM) (j) and NAA (5 μM) (h) with only partial inhibition. k Cyt D (40 μM) preventing fusion of BFA-induced PIN1-GFP intracellular aggregations. l ORY (15 μM) partial inhibition of BFA-induced PIN1-GFP intracellular aggregations, note less PIN-GFP in the perinuclear area. Scale bar = 10 μm. m Relative area of intracellular PIN1-GFP signal after 30 min of BFA treatment alone or with 30 min pre-treatment with inhibitors, auxins and cytoskeletal drugs followed by the addition of 20 μM BFA for 30 (expressed in ‰ - per mil of the total cell area). Values represent the means of the ratios between integral area of the intracellular PIN1-GFP and the total area of the cell. Error bars represent SEM from three biological repetitions, n = 3

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