Display Settings:

Items per page
We are sorry, but NCBI web applications do not support your browser and may not function properly. More information

Results: 7

1.
Figure 4

Figure 4 . From: C/EBP? regulates multiple IL-1?-induced human astrocyte inflammatory genes.

IL-1β signals through ERK1/2 and p38K to increase astrocyte BDKRB2 and COX-2 mRNA, respectively. Astrocytes were cultured for 24 h, pretreated with pathway-selective inhibitors (20 μM) and then treated with IL-1β for 12 h. Total RNA was isolated, reverse-transcribed and then assayed by RT2PCR for (A) BDKRB2 or (B) COX-2 relative transcript levels. GAPDH was used as the normalizing control; p-values are compared to control unless denoted by lines to specific comparisons.

Jerel Fields, et al. J Neuroinflammation. 2012;9:177-177.
2.
Figure 2

Figure 2 . From: C/EBP? regulates multiple IL-1?-induced human astrocyte inflammatory genes.

C/EBPβ knockdown enhances BDKRB2 and reduces COX-2 mRNA. Astrocytes were transfected with siRNA specific for C/EBPβ or siCON (100 nM), and then cultured for 24 h before being treated with IL-1β (20 ng/ml) for 12, 24 or 72 h. Total RNA was isolated, reverse-transcribed and then assayed by RT2PCR for (A) C/EBPβ, (B) BDKRB2 and (C) COX-2 relative transcripts levels. GAPDH was used as the normalizing control; p-values are compared to control unless denoted by lines to specific comparisons.

Jerel Fields, et al. J Neuroinflammation. 2012;9:177-177.
3.
Figure 6

Figure 6 . From: C/EBP? regulates multiple IL-1?-induced human astrocyte inflammatory genes.

p38K-selective small molecule inhibitor blocks IL-1β-induced COX-2 expression. (A-D) Astrocytes were cultured in 48-well plates for 24 h, treated with selective inhibitors (SB203580-20 μM, and U0126-20 μM) for 1 h and then IL-1β (20 ng/ml) for 24 h. Cells were fixed, blocked and then incubated with antibodies against GFAP (green) and COX-2 (red). All pictures were taken at ×200, and data represent at least three independent experiments in at least two independent donors.

Jerel Fields, et al. J Neuroinflammation. 2012;9:177-177.
4.
Figure 5

Figure 5 . From: C/EBP? regulates multiple IL-1?-induced human astrocyte inflammatory genes.

IL-1β signals through p38K to increase astrocyte COX-2 protein. (A-B) Astrocytes were cultured for 24 h, pretreated with pathway-selective inhibitors (SB203580-20 μM and U0126-20 μM) and then treated with IL-1β for 24 h. Total protein lysates were isolated, resolved by SDS-PAGE, transferred to a PVDF membrane and then immunoblotted for COX-2 expression. (B) COX-2 and β-actin bands were analyzed by densitometry analysis, and their intensities graphed using β-actin as the loading control; p-values are compared to control unless denoted by lines to specific comparisons. Densitometry fold change data are cumulative from two independent astrocyte donors (n = 2).

Jerel Fields, et al. J Neuroinflammation. 2012;9:177-177.
5.
Figure 3

Figure 3 . From: C/EBP? regulates multiple IL-1?-induced human astrocyte inflammatory genes.

C/EBPβ knockdown blocks IL-1β-induced C/EBPβ and COX-2 protein levels. Astrocytes were transfected with siRNA specific for C/EBPβ or siCON (100 nM) and then cultured for 24 h before being treated with IL-1β (20 ng/ml) for 24 h. (A and C) Protein lysates were isolated, resolved by SDS-PAGE, transferred to a PVDF membrane and then immunoblotted for C/EBPβ and COX-2 expression. (B and D) The densitometry graph is cumulative data from two independent astrocyte donors (n = 2) that were normalized to β-actin (B and D); p-values are compared to control unless denoted by lines to specific comparisons.

Jerel Fields, et al. J Neuroinflammation. 2012;9:177-177.
6.
Figure 1

Figure 1 . From: C/EBP? regulates multiple IL-1?-induced human astrocyte inflammatory genes.

IL-1β regulates expression of 29 of 92 human inflammation genes; C/EBPβ knockdown affects 17 of the 29 genes. Primary human astrocytes, from two independent donors, were transfected with siCON or siC/EBPβ, cultured for 24 h and then treated with IL-1β (20 ng/ml) for 12 h. Total RNA was isolated, reverse-transcribed and then assayed for human inflammation gene expression using the TaqMan® Human Inflammation Assay. Data are illustrated as a heat map; red and green represent positive and negative fold-change, respectively. Increasing color contrast corresponds to increasing absolute fold-change. Orange and blue represent positive and negative percent change, respectively, in target mRNA levels in Emphasis>/Emphasis>/EBPβ + IL-1β versus IL-1β. Each individual TaqMan® assay of the Human Inflammation Array was independently analyzed by one-way ANOVA followed by Newman-Keuls post-test to compare among control, IL-1β and Emphasis>/Emphasis>/EBPβ treatments. Significance was set at p < 0.05. (Newman-Keuls post-test: control versus IL-1β, * = p < 0.05; control versus siC/EBPβ + IL-1β, ^ = p < 0.05; siC/EBPβ + IL-1β versus IL-1β, # = p < 0.05).

Jerel Fields, et al. J Neuroinflammation. 2012;9:177-177.
7.
Figure 7

Figure 7 . From: C/EBP? regulates multiple IL-1?-induced human astrocyte inflammatory genes.

IL-1β activates astrocyte MAPK pathways, activates transcription factors and thereby regulates multiple human inflammation genes. IL-1β-mediated activation of astrocyte MAPK [p38K and ERK1/2] pathways precedes robust changes in gene expression. Multiple transcription factors traffic to astrocyte nuclei where they facilitate changes in mRNA transcription. Blocking IL-1β-mediated activation of astrocyte p38K blocks C/EBPβ translocation to nuclei [8]. The subsequent effects are similar to C/EBPβ knockdown; IL-1β-induced astrocyte TIMP-1 and COX-2 expression is blocked, and BDKRB2 expression is enhanced (Figure 4) [8]. ERK1/2 inhibition completely blocks IL-1β-induced astrocyte BDKRB2 expression and TIMP-1 expression, but enhances IL-1β-induced C/EBPβ and COX-2 expression (Figures 45 and 6) [8]. Two scenarios involving C/EBPβ could explain these events: (1) Blocking ERK1/2 signaling inhibits factors essential for IL-1β-induced astrocyte TIMP-1 expression and results in increased C/EBPβ translocation to nuclei to compensate for reduced TIMP-1 transcription. Increased C/EBPβ may account for increased COX-2. (2) Alternatively, ERK1/2 signaling may directly down-modulate IL-1β-induced astrocyte C/EBPβ; therefore, ERK1/2-selective inhibitors may increase COX-2 through an increase in CEBPβ activity. In either scenario, ERK1/2 is essential, and C/EBPβ is an auxiliary path to IL-1β-induced astrocyte TIMP-1. These data suggest MAPK, p38K and ERK1/2 are part of an intricate regulation network in which the two kinases balance one another’s activity to achieve a graded astrocyte response to CNS injury.

Jerel Fields, et al. J Neuroinflammation. 2012;9:177-177.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk