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Figure 6 | BMC Plant Biology

Figure 6

From: Iron and ferritin accumulate in separate cellular locations in Phaseolus seeds

Figure 6

Ferritin immunolocalization. Sections of P. coccineus (a, b, and f to h), P. vulgaris G14519 (i and m), and P. lunatus G25350 (j, k, l, n, o, and p) cotyledons were immunostained using antibodies raised against A. thaliana ferritin1 (AtFER1) and Alexa 546 secondary antibodies. In a, b, c, f, g, h, k, l, o, and p the seeds were soaked in water for 18-24 hours prior to PPB staining, while in i, j, m, and n dry seeds were dissected and soaked in 70% ethanol for 24 hours prior to PPB staining. b, g, i, j, k, and l: fluorescence microscopy of the immunostained tissue sections shown in a, f, m, n, o, and p respectively. a and m: Light microscopy of Perls' Prussian blue stained tissue after immunostaining. In f, n, o, and p the sections were treated with Lugol solution after immunostaining; the starch is stained black. c: Section treated like b, g, and h, but without primary antibody. h: Confocal microscopy of a single immunostained cell of P. coccineus cotyledon. d: Western blot analysis of the gel shown in e using anti-AtFER1 serum. e: Coomassie blue stained gel of 6 μg total protein from beans of P. coccineus (Pc) and P. vulgaris (Pv). M: Protein ladder, pvb: provascular bundle, am: amyloplasts, ep: epidermis, *: site where starch crystals were removed during sectioning. Scale bars in a to c and f to h: 0.1 mm, and in i to p: 0.01 mm

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