Figure 8

Effect of RNAi knock-down of Kaiso on LAMA-84. (A) Kaiso knock-down induce Wnt11 expression. LAMA-84 cells were transfected with 20nM of siRNA. Twenty-four hours later, RNA was isolated and subjected to Real Time RT-PCR to quantify expression of Wnt11, β-catenin, Kaiso and p120ctn after normalization to β-actin and compared to the scrambled knock-down cells. Columns, mean (n= 2). (B) Kaiso knock-down improve SCF expression in LAMA-84. Expression analysis of SCF gene by Real Time RT-PCR in Kaiso knock-down cells. Columns, mean (n= 2). (C) Effect of Kaiso knock-down on cell differentiation. LAMA-84 cells were transfected with 20nM of siRNA. Twenty-four hours later, RNA was isolated and subjected to Real Time RT-PCR to quantify expression of C-MyB, PU-1, C/EBPalpha and Gata-2 after normalization to β-actin and compared to the scrambled knock-down cells. Columns, mean (n= 2). (D) Knocking-down expression of Kaiso in LAMA-84 cells induced monocyte-macrophage differentiation. Flow cytometric measurement of granulocytic specific surface antigen CD15 and CD11b in LAMA-84 cells after normalization with the scrambled knock-down expression. LAMA cells were transfected with 20nM of siRNA after 50 hr-incubation. The CD117 (c-Kit receptor), CD33 and the erythroid markers GPA and CD71 were also analyzed. Columns, mean (n= 2).