Figures & data
Figure 1. I/R rats show severe pathological changes in myocardial tissues (× 400). I/R, ischemia-reperfusion.
Figure 2. CMECs were successfully cultured and identified. (a), primary CMECs of rats (7 d after adherence to wall) (× 100); (b), sub-cultured CMECs (× 100); (c), cell identification showing that factor VIII was tan-colored in cytoplasm (× 400). CMECs, cardiac microvascular endothelial cells.
Figure 3. JAZF1 plasmids were successfully transfected into CMECs. (a), relative mRNA levels of JAZF1 in response to the treatment of JAZF1 or JAZF1 shRNA; (b), relative protein levels of JAZF1 in response to the treatment of JAZF1 or JAZF1 shRNA; (c), the gray value of JAZF1 protein band in response to the treatment of JAZF1 or JAZF1 shRNA; CMECs, cardiac microvascular endothelial cells; NC, negative control; shRNA, short hairpin RNA; JAZF1, the juxtaposed with another zinc finger protein 1; *, p < 0.05 vs. the normal group; #, p < 0.05 vs. the blank, empty vector and NC shRNA group.
Figure 4. Over-expressed JAZF1 elevates levels of VEGF and Bcl-2 while reducing that of Bax, Bad, caspase-3 and caspase-9 as well as p-Bad/Bad, p-FKHR/FKHR, c-caspase3/t-caspase3 and c-caspase9/t-caspase9. (a). mRNA expression levels of related genes in cells before and after transfection; (b) & (d), protein bands of related proteins in cells of each group; (c) & (e), protein expression levels of related proteins in cells before and after transfection; NC, negative control; Akt, protein kinase (b); VEGF, vascular endothelial growth factor; Bcl-2, B-cell lymphoma-2; Bax, Bcl-2 associated X protein; Bad, Bcl-2/Bcl-XL-associated death promoter; shRNA, short hairpin RNA; JAZF1, the juxtaposed with another zinc finger protein 1; *, p < 0.05 vs. the normal group; #, p < 0.05 vs. the blank and NC groups.
Figure 5. Over-expression of JAZF1 promotes while silencing of JAZF1 inhibits cell viability. *, p < 0.05 vs. the normal group; #, p < 0.05 vs. the blank, empty vector and NC shRNA groups; NC, negative control; shRNA, short hairpin RNA.
Figure 6. More cells are arrested at the S phase after transfected with JAZF1, and few cells are arrested at the S phase after transfected with silenced JAZF1. (a), cell cycle mapping in each group; (b), the percentage of cells in each phase of cell cycle in each group; *, p < 0.05 vs. the normal group; #, p < 0.05 vs. the blank, empty vector and NC shRNA groups; NC, negative control; shRNA, short hairpin RNA; CMECs, cardiac microvascular endothelial cells; JAZF1, the juxtaposed with another zinc finger protein 1.
Figure 7. Rate of cell apoptosis of CMECs is inhibited after transfection with JAZF1, and the rate is promoted after transfection with silenced JAZF1. (a), cell apoptosis mapping measured by flow cytometry; (b), apoptosis rates in six groups; *, p < 0.05 vs. the normal group; #, p < 0.05 vs. the blank, empty vector and NC shRNA group; NC, negative control; shRNA, short hairpin RNA; CMECs, cardiac microvascular endothelial cells; JAZF1, the juxtaposed with another zinc finger protein 1.
Figure 8. Over-expressed JAZF1 promotes angiogenesis and silencing of JAZF1 inhibits angiogenesis in CMECs. (a), formation of blood vessel observed under the inverted microscope (× 100); (b), the number of blood vessel formed in each group; (c), the length of blood vessel formed in each group; *, p < 0.05 vs. the normal group; #, p < 0.05 vs. the blank, empty vector and NC shRNA group; NC, negative control; shRNA, short hairpin RNA; CMECs, cardiac microvascular endothelial cells; JAZF1, the juxtaposed with another zinc finger protein 1.
Figure 9. Over-expressed JAZF1 increases the expression of PDGF while silencing brings about the opposite results. (a), relative mRNA level of PDGF in CMECs after transfection using RT-qPCR; (b), relative protein level of PDGF in CMECs after transfection detected by western blot analysis; *, p < 0.05 vs. the normal group; #, p < 0.05 vs. the blank, empty vector and NC shRNA group; NC, negative control; shRNA, short hairpin RNA; CMECs, cardiac microvascular endothelial cells; JAZF1, the juxtaposed with another zinc finger protein 1; PDGF, platelet-derived growth factor.
Table 1. Primer sequences for RT-qPCR