Fig. 3: Sakuranetin decreases endocytosis of the PWL2 protein via the CME pathway in rice root epidermal cells.
From: Phytoalexin sakuranetin attenuates endocytosis and enhances resistance to rice blast
The root epidermal cells of rice lines LTH/35S::PWL2-GFP (a, c, e) and IRBL9-W/35S::PWL2-GFP (b, d, f) treated with 25 μM BFA for 90 min. g Quantification of the relative size of the BFA bodies shown in the images a–f (nLTH/35S::PWL2-GFP #19 = 63; nLTH/35S::PWL2-GFP #21 = 76; nLTH/35S::PWL2-GFP #22 = 56; nIRBL9-W/35S::PWL2-GFP #4 = 71; nIRBL9-W/35S::PWL2-GFP #9 = 44; nIRBL9-W/35S::PWL2-GFP #19 = 38). The root epidermal cells of seedlings of the rice line LTH/35S::PWL2-GFP were co-treated for 90 min with 100 µM SAK and 25 µM BFA (i), or with 25 µM BFA and an equivalent volume of DMSO as a control (h). j Quantification of the relative sizes of the BFA bodies shown in the images h, i (nDMSO = 15; nSAK = 17). The root epidermal cells of rice NPB/ProOsPIN1b::OsPIN1b-GFP (#11) (k, l) and NPB/ProOsPIN2::OsPIN2-GFP (#17) (m, n) co-treated for 90 min with either 100 μM SAK, 25 μM BFA and 50 μM TyrA23 (l, n), or with 25 μM BFA, 50 μM TyrA23 and an equivalent volume of DMSO as a control (k, m). o Quantification of the relative size of the BFA bodies shown in the images k–n (DMSO: nNPB/ProOsPIN1b::OsPIN1b-GFP = 48; nNPB/ProOsPIN2::OsPIN2-GFP = 16; SAK: nNPB/ProOsPIN1b::OsPIN1b-GFP = 92; nNPB/ProOsPIN2::OsPIN2-GFP = 39). The root epidermal cells of rice LTH/35S::PWL2-GFP were co-treated for 90 min with 25 μM BFA and DMSO as a control (p), or with 50 µM TyrA23 and 25 μM BFA (q), or with 100 μM SAK, 50 µM TyrA23 and 25 μM BFA (r). s Quantification of the relative sizes of the BFA bodies shown in the images p–r (n = 143 in image p; n = 80 in image q; n = 188 in image r). SAK sakuranetin, TyrA23 tyrphostin A23. Data are means ± SE; P values were generated with independent-samples two-sided Student’s t-test in images g, j, o and s. Scale bar = 20 μm in images a–f, h and i, Scale bar = 10 μm in images k–n and p–r). Arrowheads indicate the BFA bodies.
