il 1β production  (ATCC)

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E expression is elevated by interleukin 1 and other interleukin 1-induced factors

doi: 10.1186/1742-2094-8-175

Figure Lengend Snippet: RT-PCR primers

Article Snippet: Rat recombinant mature IL-1β (IL-1β holoprotein cleavage product) was purchased from Sigma (St. Louis MO), secreted APP (sAPP) was purified from a recombinant expression system as described previously [ ], and L-glutamate was from Sigma (St. Louis MO).

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E expression is elevated by interleukin 1 and other interleukin 1-induced factors

doi: 10.1186/1742-2094-8-175

Figure Lengend Snippet: IL-1β induces ApoE expression in vivo . Proteins were extracted from the left cerebral cortex of unoperated rats ( con ), of rats implanted in the right cerebrum with a pellet containing vehicle alone ( sham ), and of rats implanted in the right cerebrum with a slow-release pellet containing 100 ng IL-1β ( IL-1β pellet ). A: RNA was extracted from the left cerebral cortex of con (n = 6), sham (n = 5), and IL-1β pellet (n = 7) rats and subjected to gel-based RT-PCR for ApoE and GAPDH. B: Quantification of the ApoE mRNA (relative to that for GAPDH) by densitometry of the RT-PCR image or by quantitative (real-time) RT-PCR ( qRT-PCR ); values reflect the percent of control as mean ± SEM. C: Proteins were extracted from con (n = 3), sham (n = 3), and IL-1β pellet (n = 8) rats and subjected to western immunoblot analysis for ApoE. D: Quantification of autoradiography; values reflect percent of control as mean ± SEM. * p ≤0.05; ** p < 0.01 versus control.

Article Snippet: Rat recombinant mature IL-1β (IL-1β holoprotein cleavage product) was purchased from Sigma (St. Louis MO), secreted APP (sAPP) was purified from a recombinant expression system as described previously [ ], and L-glutamate was from Sigma (St. Louis MO).

Techniques: Expressing, In Vivo, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Autoradiography

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E expression is elevated by interleukin 1 and other interleukin 1-induced factors

doi: 10.1186/1742-2094-8-175

Figure Lengend Snippet: IL-1β induces increases in mRNAs encoding ApoE, βAPP, and the proinflammatory factors IL-1β, ICE, IL-1α, and TNF . RNA was extracted from the left cerebral cortex of unoperated rats ( Control ), rats implanted in the right cerebrum with a pellet containing vehicle alone ( Sham ), and rats implanted in the right cerebrum with a slow-release pellet containing 100 ng IL-1β ( IL-1β Pellet ); A: semi-quantitative RT-PCR was performed on the RNA. B: Results quantified by densitometry showed significant differences in mRNA levels from brains with IL-1β pellets compared to those with sham pellets, or from those serving as controls; p ≤0.05 for each gene product.

Article Snippet: Rat recombinant mature IL-1β (IL-1β holoprotein cleavage product) was purchased from Sigma (St. Louis MO), secreted APP (sAPP) was purified from a recombinant expression system as described previously [ ], and L-glutamate was from Sigma (St. Louis MO).

Techniques: Quantitative RT-PCR

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E expression is elevated by interleukin 1 and other interleukin 1-induced factors

doi: 10.1186/1742-2094-8-175

Figure Lengend Snippet: IL-1β induces expression of IL-1α, ApoE, and βAPP . Immunofluorescence was performed on tissue sections from the neocortex or hippocampus (CA1) of unoperated rats (n = 4) ( Control ), from rats implanted with a pellet containing vehicle alone (n = 6) ( Sham ), and from rats implanted with a slow-release pellet containing 100 ng IL-1β (n = 6) ( IL-1β Pellet ). Primary antibodies were A: ApoE, B: IL-1α, or C: βAPP; the red immunofluorescence signal is countered by a blue DAPI stain of cell nuclei. Immunofluorescence was quantified as described in Materials and Methods, and is expressed as mean ± SEM. All treatment conditions were significantly different from each other except with regard to IL-1α, where only the comparison of IL-1β pellet to the other two treatments was significant (* p ≤0.05).

Article Snippet: Rat recombinant mature IL-1β (IL-1β holoprotein cleavage product) was purchased from Sigma (St. Louis MO), secreted APP (sAPP) was purified from a recombinant expression system as described previously [ ], and L-glutamate was from Sigma (St. Louis MO).

Techniques: Expressing, Immunofluorescence, Staining

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E expression is elevated by interleukin 1 and other interleukin 1-induced factors

doi: 10.1186/1742-2094-8-175

Figure Lengend Snippet: Elevation of ApoE expression in neuronal cells by stimuli relevant to Alzheimer's disease . Primary cortical neurons or NT2 cells were treated with IL-1β (30 ng/ml), Aβ 1-42 (10 μM), sAPP (30 nM), or Glu (50 μM) for 20 h. A: RNA was extracted from the cultures for RT-PCR of ApoE and GAPDH as illustrated. The results were quantified by densitometry, and values represent the mean ± SEM for the ApoE amplimer relative to that for GAPDH. B: Cell lysates from the cultures were equilibrated for total protein levels and analyzed for ApoE content by western blot, also quantified by densitometry (* p < 0.05, ** p < 0.01).

Article Snippet: Rat recombinant mature IL-1β (IL-1β holoprotein cleavage product) was purchased from Sigma (St. Louis MO), secreted APP (sAPP) was purified from a recombinant expression system as described previously [ ], and L-glutamate was from Sigma (St. Louis MO).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E expression is elevated by interleukin 1 and other interleukin 1-induced factors

doi: 10.1186/1742-2094-8-175

Figure Lengend Snippet: Potential for indirect effects of IL-1β . A: Primary cortical neurons were treated for 20 h with the indicated concentrations of IL-1β, and glutamate levels were assessed in the medium (* p < 0.05). B: Primary cortical neurons were treated for 20 h with the indicated concentrations of IL-1β, and sAPP levels were assessed in concentrated aliquots of the culture medium by western blot; * p < 0.05. C: Primary cortical neurons were treated for 20 h with glutamate (50 μM) and Aβ 1-42 (10 μM) alone, or with a combination of the two; immunofluorescence was performed for ApoE (green) and βAPP (red). D: The bar graph shows integrated fluorescence intensity quantified for the ApoE signal in the cultures shown in panel C; n = 3 cultures for each treatment (* p < 0.05 compared to the untreated sample).

Article Snippet: Rat recombinant mature IL-1β (IL-1β holoprotein cleavage product) was purchased from Sigma (St. Louis MO), secreted APP (sAPP) was purified from a recombinant expression system as described previously [ ], and L-glutamate was from Sigma (St. Louis MO).

Techniques: Western Blot, Immunofluorescence, Fluorescence

Journal: Journal of Neuroinflammation

Article Title: Apolipoprotein E expression is elevated by interleukin 1 and other interleukin 1-induced factors

doi: 10.1186/1742-2094-8-175

Figure Lengend Snippet: Roles for p38-MAPK, ERK, and JNK in elevation of ApoE expression by IL-1β, Aβ 1-42 , sAPP, and glutamate . Cultures of primary cortical neurons or NT2 cells were treated for 20 h with A: IL-1β (30 ng/ml), B: Aβ 1-42 (10 μM), C: sAPP (30 nM), D: or glutamate (50 μM). In parallel cultures, these agents were preceded by 5 μM of an inhibitor of p38-MAPK (SB), the ERK pathway (U0126), or JNK (JNKin). Lysates were examined by western blot analysis of full-length ApoE, and autoradiographs were quantified by densitometry (bar graphs). Values represent mean ± SEM (n = 4); *p < 0.05, **p < 0.01.

Article Snippet: Rat recombinant mature IL-1β (IL-1β holoprotein cleavage product) was purchased from Sigma (St. Louis MO), secreted APP (sAPP) was purified from a recombinant expression system as described previously [ ], and L-glutamate was from Sigma (St. Louis MO).

Techniques: Expressing, Western Blot