Table 4.

Effects of actin network remodelling and cytosolic Ca ²⁺ plumes on vesicle distribution in trans-differentiating adaxial epidermal cells of culturedV. faba cotyledons.

Cotyledons were freshly harvested (0 h, control), or cultured for 15 h in the absence/presence of an endo- and exocytosis inhibitor, Brefeldin A (BFA), an actin depolymerization drug, latrunculin B, an actin stabilization drug, jasplakinolide, or a DHP-receptor Ca²⁺-channel blocker, nifedipine. Thereafter, transverse sections of treated cotyledons were stained with the membrane dye FM4-64FX for 10 min. Fluorescence was measured as total pixel intensities in specified cell regions. These values were adjusted for fluorescence from FM4-64FX located in the plasma membrane using the BFA values (for more details, see Supplementary Table S1) to provide estimates of cytoplasmic fluorescence. Data are the mean differences ±SE (n=4) between mean pixel intensities of 25 cells measured per cotyledon recorded across four replicate cotyledons for the specified treatment minus fluorescence derived from a similar population of cells exposed to BFA. See Supplementary Table S1 for procedure used to estimate the SE of the difference between two means of unpaired observations.

Treatment	Fluorescence intensity (arbitrary units)
	Outer periclinal cytoplasm	Anticlinal cytoplasm	Inner periclinal cytoplasm	Total
0 h Control	43 ± 7	0	33 ± 6	76 ± 10
15 h Control	385 ± 17	25 ± 5	13 ± 4	424 ± 19
15 h Latrunculin B (100 nM)	184 ± 18	121 ± 13	180 ± 15	505 ± 27
15 h Jasplakinolide (100 nM)	184 ± 25	101 ± 10	176 ± 25	461 ± 37
15 h Nifedipine (100 μM)	165 ± 11	111 ± 7	142 ± 8	418 ± 15