Research Paper Volume 11, Issue 10 pp 3376—3391

Figure 4. ID4 rescued the pro-leukemia effects of miR-335 in leukemic cell-lines. (A–B) ID4 mRNA expression affected by miR-335 transfection. ID4 mRNA expression was significantly reduced after miR-335 overexpression in both K562 and HL60 cell-lines. (C–D) Confirmation of ID4 mRNA expression after ID4 restoration. ID4 mRNA expression was significantly upregulated after ID4 transfection in both K562/miR-335 and HL60/miR-335 cells. (E) ID4 protein expression affected by miR-335 overexpression and ID4 restoration. ID4 protein expression was significantly reduced after miR-335 overexpression in both K562 and HL60 cell-lines, and was increased after ID4 restoration. (F–G) The effect of ID4 restoration on cell proliferation. Restoration of ID4 significantly reduced the proliferation ability in K562/miR-335 and HL60/miR-335 cells. (H–I) The effect of ID4 restoration on cell apoptosis. Restoration of ID4 significantly increased the apoptosis ratio in K562/miR-335 and HL60/miR-335 cells. (J) The expression of proliferation-related proteins (PCNA and Cyclin D1) and apoptosis-related proteins (Caspase-3) affected by ID4 restoration. The expression of proliferation-related proteins (PCNA and Cyclin D1) was decreased, whereas the apoptosis-related proteins (Caspase-3) expression was increased after ID4 restoration in K562/miR-335 and HL60/miR-335 cells. *, P<0.05; **, P<0.01; ***, P<0.001.