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Development of a novel chronic kidney disease mouse model to evaluate the progression of hyperphosphatemia and associated mineral bone disease Takashi Tani 1, 2 , Hideo Orimo 2 , Akira Shimizu 3 , Shuichi Tsuruoka 1 Supplementary Figures: Supplementary Figure S1. Micrographs of H&E, EMG, Von kossa and Alizarin Red stained sections of the thoracic aorta from A6P2 and A6P4 mice. Sections of the thoracic aorta for A6P2 (a-d) and A6P4 (e-h) groups are shown. Medial arterial calcification was observed, as Von kossa and Alizarin Red staining positive lesion, in part of A6P2 and A6P4 groups` mice (c,d,g,h, black arrow). Scale bar, 500 μm; H&E, hematoxylin and eosin staining; EMG, elastica Masson-Goldner staining. x200 HE Von-Kossa EMG Alizarin Red e f b A6P4 c g d A6P2 h a HE

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Developmentofanovelchronickidneydiseasemousemodeltoevaluatethe progression of hyperphosphatemia and associated mineral bonediseaseTakashiTani1,2,HideoOrimo2,AkiraShimizu3,ShuichiTsuruoka1

SupplementaryFigures:

Supplementary Figure S1. Micrographs of H&E, EMG, Von kossa andAlizarinRedstainedsectionsof the thoracicaorta fromA6P2andA6P4mice.SectionsofthethoracicaortaforA6P2(a-d)andA6P4(e-h)groupsare shown.Medial arterial calcification was observed, as Von kossa andAlizarinRedstainingpositivelesion,inpartofA6P2andA6P4groups`mice(c,d,g,h, black arrow). Scale bar, 500 μm; H&E, hematoxylin and eosinstaining;EMG,elasticaMasson-Goldnerstaining.

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SupplementaryFigureS2.Computedtomographyimagingoftheidenticalmouse between 16 and 20 weeks of age. Computed tomography (CT)imagingwasperformedrepeatedly for identicalmouseeveryotherweekbetween14and20weeksofage.CTimageswerereconstructedassagittalplaneimages(a,c,e)andcoronalplaneimages(b,d,f).Vascularcalcificationbecame prominent at 16weeks of age, i.e. two-weeks after phosphorusloading (a, b, arrows). Calcificationbecame severer as a time-dependentmannerat18weeksofage(c,d,arrows)and20weeksofage(e,f,arrows).

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