Supplemental Materials

Figure Legends

Suppl. Figure 1. Time course of LV function and apoptosis after chronic pressure overload with/without caspase inhibitor (CI). There was no significant difference in LVEF(%) and apoptosis (%) at 1wk after transverse aortic constriction (TAC) between CI and vehicle (veh) group. At 3wk after TAC, LVEF(%) was preserved in CI group compared to veh. Non-myocyte apoptosis was significantly reduced in CI group compared to veh at 3wk after TAC. Myocyte apoptosis was increased at 3wk after TAC with no significant difference between CI and veh group. LV ejection fraction; LVEF(%), non-myocyte; NM (diamond), myocyte; Myo (circle).

Suppl. Figure 2. (A) The correlation of TUNEL-positive cells and cleaved caspase-3 (CC3) positive cells in chronic pressure overload. (B) Representative pictures of immunostaining with rhodamine-conjugated wheat germ agglutinine (WGA) (upper panel) and CC3 (lower panel) with DAPI; WGA (membrane, red), CC3 (apoptosis, green), DAPI (nucleus, blue). The yellow arrow in lower panel indicates CC3-positive myocyte.

Suppl. Figure 3 Representative photographs of wheat germ agglutinin (WGA) staining for measuring myocyte cross-sectional area. The outlines of myocytes were shown by WGA staining (white). Caspase inhibition (CI), vehicle (veh), transverse aortic constriction (TAC).

Suppl. Figure 4. Fibrosis after chronic pressure overload with/without caspase inhibition. Fibrosis was determined by three different methods. Representative photographs of fibrosis measurement from (A) picrosirius red (PSR)-staining; fibrosis (red), (B) Masson’s trichrome staining; collagen (bluish purple), and (C) dual immunohistochemical staining with connective tissue growth factor (CTGF) (green) and wheat germ agglutinin (WGA) (red). Caspase inhibition (CI), vehicle (veh), transverse aortic constriction (TAC).

Suppl. Figure 5. Endothelial cells in vitro tube formation assay for measuring angiogenesis. (A) Caspase inhibition (1μM) shows trends to increase of tube formation of HMVEC-Cs (human cardiac microvascular endothelial cells), and further increase with phenylephrine (100μM). (B) Representative pictures for tube formation of HMVEC-Cs on matrigel. Caspase inhibition (CI), vehicle (veh), phenylephrine (PE).

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