MMP3 regulate the expression of Nox1 in 6-OHDA treated N27 cells.

<p>(A) A representative RT-PCR performed on various Nox subunits from RNA preparation of N27 cells treated with 100 µM 6-OHDA. (B) A representative RT-PCR performed on MMP3 and Nox1 from RNA preparation of N27 cells. N27 cells were transfected with two different siRNAs against MMP-3 and Nox1 and their scrambled sequences (scr-siRNA) and subsequently treated with 6-OHDA for 6 h. Densitometric analysis of the RT-PCR results normalized against GAPDH and expressed as % induction compared with respective untreated control. * p<0.05, ** p<0.01, ***p <0.001 vs. control ; ###p<0.001 vs. 6-OHDA control. (C) Representative photographs of western blot for MMP3 and Nox1 on lysate of N27 cells transfected with two different siRNAs against MMP-3 and Nox1 and scr-siRNA and subsequently treated with 6-OHDA for 24 h. Densitometric analysis of the Western blot results normalized against β-actin and expressed as % induction compared with respective untreated control. ** p<0.01 vs. scr-siRNA control ; ##p<0.01 vs. scr-siRNA+6-OHDA control. (D) A representative western blot against Nox1 on lysate of N27 cells pre-treated with 1 µM and 5 µM N-isobutyl-N-(4-methoxyphenylsulfonyl) glycyl hydroxamic acid (NNGH) for 30 min followed by exposure to 100 µM 6-OHDA for 24 h. Densitometric analysis of the western blots results normalized against β-actin and expressed as % expression compared with respective untreated control. ***p<0.001 vs. control ; ##p<0.01vs. 6-OHDA control. Results are presented as the mean ± SEM, n = 8. The whole experiment has been repeated four times with similar results.