Fig. 3: Systemic endocytosis induced by La(III) was dependent on AtrbohD.

a The representative CLSM images of Col-0, DPI-treated Col-0 (leaves or stems treated with DPI), atrbohD, and grafted atrbohD (shoots from atrbohD were grafted onto Col-0 roots and shoots from Col-0 were grafted onto atrbohD roots) root cells. Leaves of all of these Arabidopsis were treated with 0, 30, or 80 µM La(III) for 12 h and then roots were stained with FM4-64. Bar = 10 μm. Representative images from six independent measurements and three biological replicates (each replicate represents an independently treated plant) per measurement are represented. b Quantitative analysis of the average fluorescent area of FM4-64 in Col-0, Col-0 leaves or stems treated with DPI, atrbohD, and grafted atrbohD (shoots from atrbohD were grafted onto Col-0 roots and shoots from Col-0 were grafted onto atrbohD roots) root cells with endocytosis (data come from a). Values shown are means ± SEM, one-way ANOVA analysis with LSD multiple comparisons test (n = 6, **p < 0.01). c, d The expression levels of AtrbohD and AtrbohF transcripts in Col-0 leaves and roots 12 h after treatment of leaves with 0, 30, or 80 μM La(III). The expression levels were quantified by using quantitative RT-PCR. ACTIN2 (c) and EF1α (d) were as two references. Six independent measurements and three replicates per measurement were conducted. Values shown are means ± SEM, one-way ANOVA analysis with LSD multiple comparisons test (n = 6, *p < 0.05, **p < 0.01, n.s.: no significance).