Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Remifentanil upregulates hepatic IL-18 binding protein (IL-18BP) expression through transcriptional control.

doi: 10.1038/s41374-018-0111-y

Figure Lengend Snippet: Fig. 1 Remifentanil increased the IL-18BP expression level in normal rat livers. Western blot (a) and QPCR (b) analyses showed that intravenous remifentanil treatment significantly increased the hepatic IL-18BP levels at 6 h and further induced a three-fold induction at 24 h in normal rats; n = 5 in each group, with a representative western blot shown. The data are expressed as the means ± SD. *P < 0.05, at the indicated times, vs. the Control group (Con). c Immunohistochemical staining of liver samples from the control and 24-h treatment groups showed remifentanil-induced IL- 18BP was mainly expressed by hepatocytes. Original magnification in C, ×40

Article Snippet: To assess the expression of IL-18BP, paraffin-embedded liver sections were deparaffinized, blocked in 3% albumin from bovine serum in PBS for 30 min, and incubated with an antibody against IL-18BP (1:50, sc-9464; Santa Cruz) at 4 °C overnight followed by horseradish peroxidase-linked anti-goat IgG (Santa Cruz).

Techniques: Expressing, Western Blot, Control, Immunohistochemical staining, Staining

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Remifentanil upregulates hepatic IL-18 binding protein (IL-18BP) expression through transcriptional control.

doi: 10.1038/s41374-018-0111-y

Figure Lengend Snippet: Fig. 2 Remifentanil upregulated IL-18BP mRNA expression in hepatocytes. QPCR analyses showed significantly increased expression of IL-18BP mRNA levels 6 and 24 h after treatment with 100 ng ml−1 of remifentanil (R) in a mouse primary hepatocytes, b LO2 cells, and c Hep1-6 cells. d Naloxone (Nal) did not affect the effects of remifentanil on IL-18BP mRNA expression in LO2 cells. e Western blot analyses showed that preexposure of LO2 cells to remifentanil for 24 h significantly inhibited IL-18- activated p65 phosphorylation. The inhibition was absent when the IL-18BP genes in LO2 cells were silenced with IL-18BP siRNA. n = 3 in each group in a, c, d, e. n = 5 in each group in b. The data are expressed as the means ± SD. *P < 0.05 vs. the Control group (Con). #P < 0.05 vs. the IL-18-treated group. &P < 0.05 vs. the IL-18+R group

Article Snippet: To assess the expression of IL-18BP, paraffin-embedded liver sections were deparaffinized, blocked in 3% albumin from bovine serum in PBS for 30 min, and incubated with an antibody against IL-18BP (1:50, sc-9464; Santa Cruz) at 4 °C overnight followed by horseradish peroxidase-linked anti-goat IgG (Santa Cruz).

Techniques: Expressing, Western Blot, Phospho-proteomics, Inhibition, Control

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Remifentanil upregulates hepatic IL-18 binding protein (IL-18BP) expression through transcriptional control.

doi: 10.1038/s41374-018-0111-y

Figure Lengend Snippet: Fig. 7 Schematic diagram of the effects of remifentanil on hepatic IL- 18BP expression and IL-18 dysfunction-mediated liver injury. Remi- fentanil significantly increases phosphorylation of transcription factors STAT1 and C/EBP β at the IL-18BP gene promoter region, therefore, transcriptionally activates the IL-18BP gene and upregulates the expression of IL-18BP protein in hepatocytes. Remifentanil-induced IL-18BP significantly inhibits IL-18 signaling in livers and alleviates IL-18 dysfunction-mediated liver injury

Article Snippet: To assess the expression of IL-18BP, paraffin-embedded liver sections were deparaffinized, blocked in 3% albumin from bovine serum in PBS for 30 min, and incubated with an antibody against IL-18BP (1:50, sc-9464; Santa Cruz) at 4 °C overnight followed by horseradish peroxidase-linked anti-goat IgG (Santa Cruz).

Techniques: Expressing, Phospho-proteomics

Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Remifentanil upregulates hepatic IL-18 binding protein (IL-18BP) expression through transcriptional control.

doi: 10.1038/s41374-018-0111-y

Figure Lengend Snippet: Fig. 6 Suppression of STAT1 or C/EBP β inhibited remifentanil- induced IL-18BP mRNA expression. LO2 cells were transfected with STAT1 or C/EBP β-targeting siRNAs before exposure to 100 ng ml−1

Article Snippet: To assess the expression of IL-18BP, paraffin-embedded liver sections were deparaffinized, blocked in 3% albumin from bovine serum in PBS for 30 min, and incubated with an antibody against IL-18BP (1:50, sc-9464; Santa Cruz) at 4 °C overnight followed by horseradish peroxidase-linked anti-goat IgG (Santa Cruz).

Techniques: Expressing, Transfection