Notch inhibition affects ECM remodelling gene expression and inflammatory processes (A) ISH for ctssb.1 and mmp9, revealing high expression at the inner injury border at 36 hpci. (B) ISH against tnfrsf9a on sections from an injured ventricle at 3 dpci, showing gene expression in cells with endocardial morphology within the injury site. (C) mpeg1- FISH combined with IHC, showing mpeg+ cells (yellow arrowheads) in contact with GFP+ cells. mpeg1+ cell accumulations at the injury site (is) locally coincides with high endocardial cell abundance (brackets). (D) ISH on consecutive sections for l-plastin showing high l-plastin expression in the injury site at 36 hpci. (E) ISH against mmp9 (3 dpci), showing high numbers of positive cells within the injury site (is) of RO-treated hearts. (F) ISH against ctssb.1 (7 dpci), showing high numbers of positive cells within the injury site (is) of RO-treated hearts. (G) qPCR analysis showing increased ctssb.1 mRNA levels in injured ventricles at 7 dpci after RO-treatment. (mean± s.d., t-test, *P<0.05. (H) qPCR analysis of tnfrsf9a expression in ventricles at 7 dpci after treatment with RO or DMSO mean± s.d., t-test, *P<0.05). (I) FISH against l-plastin combined with IHC against GFP and MF20 on sections of ET33-mi60a transgenic hearts (7 dpci) after RO- or DMSO-treatment. ROtreatment results in an increased abundance of l-plastin+ macrophages associated to the endocardium. Dotted lines delineate the injury site (is). Scale bars: 100 μm.
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