Journal: Journal of Neuroinflammation

Article Title: All- trans retinoic acid induces COX-2 and prostaglandin E 2 synthesis in SH-SY5Y human neuroblastoma cells: involvement of retinoic acid receptors and extracellular-regulated kinase 1/2

doi: 10.1186/1742-2094-4-1

Figure Lengend Snippet: ATRA up-regulates COX-2 expression and increases PGE 2 in SH-SY5Y human neuroblastoma cells . (a,b) Expression of COX-1 and COX-2 proteins was analyzed by western blot in SH-SY5Y human neuroblastoma cells incubated for 24 h with the indicated concentrations of ATRA or incubated with 10 μM ATRA for the indicated times. (c) For comparison, expression of COX-2 protein was also analyzed in cells incubated with the inflammatory cytokine interleukin-1 β (IL-1β,10 ng/ml). (d) Expression of COX-2 mRNA, analyzed by semiquantitative RT-PCR in cells incubated for 24 h with 10 μM ATRA or 10 ng/ml IL-1β. (e) PGE 2 production in SH-SY5Y human neuroblastoma cells incubated for 24 h with 10 μM ATRA or 10 ng/ml IL-1β. PGE 2 in the medium was determined in triplicate in four separate experiments (for statistical purposes n = 4; * P < 0.01 vs control). (a,b,c,d) Each photograph represents at least three repeated experiments. Equal protein or mRNA loading were confirmed by probing with an anti-α-actin antibody or by co-amplification of 18 S RNA, respectively Normalized density ratio of COX-2 over either α-actin or 18 S RNA is indicated for each band.

Article Snippet: The SH-SY5Y human neuroblastoma cell line (N-type cells, derived from the parental cell line SK-N-SH; Biedler et al. 1973) was obtained from American Type Culture Collection (Cat #: CRL-2266; ATCC, Manasas, VA).

Techniques: Expressing, Western Blot, Incubation, Comparison, Reverse Transcription Polymerase Chain Reaction, Control, Amplification

Journal: Journal of Neuroinflammation

Article Title: All- trans retinoic acid induces COX-2 and prostaglandin E 2 synthesis in SH-SY5Y human neuroblastoma cells: involvement of retinoic acid receptors and extracellular-regulated kinase 1/2

doi: 10.1186/1742-2094-4-1

Figure Lengend Snippet: ATRA-induced COX-2 protein expression and PGE 2 production are inhibited by RAR pan-antagonist LE540 or MEK-1 inhibitor PD98059 . (a) Western blot analysis of the expression of COX-2 protein in SH-SY5Y human neuroblastoma cells pre-incubated for 1 h with either RAR pan-antagonist LE540 2.5 μM (left) or 50 μM MEK-1 inhibitor PD98059 (right) and then incubated with ATRA (10 μM, 24 h). Equal protein loading was confirmed by probing with an anti-α-actin antibody. Normalized density ratio of COX-2 over α-actin is indicated for each band. Each experiment represents at least three repeated experiments. (b) PGE 2 production in SH-SY5Y human neuroblastoma cells incubated for 24 hours with ATRA 10 μM in cells and pre-treated for 1 hour with either RAR pan-antagonist LE540 2.5 μM or 50 μM MEK-1 inhibitor PD98059. PGE 2 in the medium was determined in triplicate in four separate experiments (for statistical purposes n = 4) (*P < 0.01 vs other groups)

Article Snippet: The SH-SY5Y human neuroblastoma cell line (N-type cells, derived from the parental cell line SK-N-SH; Biedler et al. 1973) was obtained from American Type Culture Collection (Cat #: CRL-2266; ATCC, Manasas, VA).

Techniques: Expressing, Western Blot, Incubation

Journal: Journal of Neuroinflammation

Article Title: All- trans retinoic acid induces COX-2 and prostaglandin E 2 synthesis in SH-SY5Y human neuroblastoma cells: involvement of retinoic acid receptors and extracellular-regulated kinase 1/2

doi: 10.1186/1742-2094-4-1

Figure Lengend Snippet: ATRA increases the activity of the human COX-2 promoter and its effect is inhibited by RAR-pan-antagonist LE540 or MEK-1 inhibitor PD98059 . (a) Schematic of the COX-2 human promoter construct phPES2 containing the promoter fragment -327 to +59.. (b) ATRA (10 μM, 24 h) increases the activity of the human COX-2 gene promoter transfected in SH-SY5Y human neuroblastoma cells. For comparison, the effect of IL-1β (10 ng/ml, 24 h) is also shown. COX-2 promoter activity was determined in triplicate in four separate experiments (for statistical purposes n = 4; *P < 0.01 vs other groups; **P < 0.01 vs control). (c) ATRA (10 μM, 24 h) increases the expression of RAR-β (left) and induces ERK1/2 phosphorylation (right). Normalized density ratio of either RAR-β or ERK1/2 over α-actin is indicated for each band. Each photograph represents at least three repeated experiments. (d) Inhibition of ATRA-induced COX-2 promoter activity by the RAR-pan-antagonist LE540 and the MEK-1 inhibitor PD98059. Transiently transfected cells were pre-incubated for 1 h with either 2.5 μM LE540 or 50 μM PD98059 and then with ATRA (10 μM, 24 h). COX-2 promoter activity was measured in triplicate in four separate experiments (for statistical purposes n = 4) (*P < 0.01 vs other groups)

Article Snippet: The SH-SY5Y human neuroblastoma cell line (N-type cells, derived from the parental cell line SK-N-SH; Biedler et al. 1973) was obtained from American Type Culture Collection (Cat #: CRL-2266; ATCC, Manasas, VA).

Techniques: Activity Assay, Construct, Transfection, Comparison, Control, Expressing, Phospho-proteomics, Inhibition, Incubation