Differentiation of transfected HL60 cells exposed to 1,25D or ATRA.

<p>HL60 cells were transfected with plasmids containing either control shRNA (CtrA) or <i>POR</i> shRNA and then were exposed to either 1,25D (10 nM) or ATRA (1 μM) for 96 h. The expression of differentiation markers CD11b (upper graph) and CD14 (bottom graph) was detected using flow cytometry. Mean values (±SEM) of percentages of positive cells are presented in Y-axis. Results that differ significantly from the result for respective control cells are marked with asterisks (*p<0.05; ***p<0.001), while the sample which differs from the appropriate CtrA sample is marked with hash (^#p<0.05).</p

Responses of AML patient's mononuclear cells to 1,25D or ATRA.

<p>Cells from peripheral blood of AML patients were exposed for 96 h to 10 nM 1,25D or to 1 μM ATRA. Then the expression of CD14 antigen in 1,25D-treated blast cells was studied in flow cytometry. From remaining 1,25D-treated blasts and from ATRA-treated blasts mRNA was isolated and the expression of <i>CYP24A1</i> and <i>POR</i> genes was studied in Real Time PCR.</p

Expression of <i>POR</i> gene and protein in transfected HL60 cells.

<p>HL60 cells were transfected with plasmids containing either control shRNA (CtrA) or <i>POR</i> shRNA and then (A) the expression of <i>POR</i> mRNA was tested in Real Time PCR. The bar charts show mean values (±SEM) of fold changes in mRNA levels relative to <i>GAPDH</i> mRNA levels. The control sample was calculated as 1. (B) The expression of POR protein in mitochondria of the cells exposed to 1,25D (10 nM) or ATRA (1 μM) for 96 h was detected using Western blot. Mitochondrial proteins were analyzed using anti-POR and anti-actin antibodies. Optical densities (OD) of POR bands divided by ODs of respective actin bands were calculated from 4 blots and mean values are given below the blots.</p

Responses of mononuclear cells from healthy donors to 1,25D or ATRA.

<p>Cells from peripheral blood of healthy donors were exposed for 96 h to 10 nM 1,25D or to 1 μM ATRA. Then the mRNA was isolated and the expression of <i>CYP24A1</i> and <i>POR</i> genes was studied in Real Time PCR.</p