Supplementary material
Supplementary material
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- Figure S1 - Expression of 12-LOX in NCI-H292. Cells transfected with 12R-LOX siRNAs (si-12R-LOX), 12S-LOX siRNAS (si-12S-LOX) or control si-RNAs (si-ctrl) (100 nM, final concentration) were stimulated with PMA (20 nM) for 24 h. Then, total RNA or cell lysates were obtained. Real time RT-PCR was performed using specific primers for 12S-LOX (panel A) or 12R-LOX (panel B). In C) western blots from non-stimulated NCI-H292 cells transfected with si-ctrl or si-12R-LOX were performed using anti-12R-LOX antibodies, followed by stripping and re-probing of membranes with anti-β-actin (ACTB) antibodies. Jurkat lane, corresponds to a cell extract from Jurkat clone E6-1 cell line (positive control for the antibody). The arrow indicates 12R-LOX specific band. Asterisk points non-specific band detected by the antibody. Control siRNAs did not modify 12-LOX expression neither in basal or PMA-induced conditions (data not shown). D) Quantification of 12R-LOX band intensity normalised to ACTB from C). In A) and B), Mean ± SD of triplicates from three independent experiment is shown. * p<0.05 vs. cells transfected with control siRNAs (si-ctrl).