Journal: PLoS ONE

Article Title: IL-6-Mediated Activation of Stat3α Prevents Trauma/Hemorrhagic Shock-Induced Liver Inflammation

doi: 10.1371/journal.pone.0021449

Figure Lengend Snippet: Rats were subjected to sham protocol (Sham, n = 3), unresuscitated T/HS (UHS, n = 3), T/HS treated with placebo at the beginning of resuscitation (SBR50, n = 4), T/HS treated with IL-6 at the beginning of resuscitation (SBR50/IL-6, n = 4), T/HS preceded by treatment with GQ-oligodeoxynucleotide (GQ-ODN) 24 hours prior to resuscitation with IL-6 (SBR50/IL-6/G, n = 3), or T/HS preceded by treatment with nonspecific-ODN (NS-ODN) 24 hours prior to resuscitation with IL-6 (SBR50/IL-6N, n = 3). The livers were harvested 60 minutes after the start of resuscitation. MPO staining was performed in paraformaldehyde-fixed livers. Representative photomicrographs of 1000x fields of liver specimens from each experimental group are shown (2A). Arrows indicate MPO-positive cells. In , MPO-positive cells were counted in 20 hpf; data presented are the means ± SEM of each group (n≥5). Bars marked with an asterisk (*) differ significantly within the pair (p<0.05). shows representative immunoblots of protein extracts of whole liver from each group, a negative control (NC) cells and positive control (PC) cells developed with monoclonal antibodies to either phosphorylated (p)Stat3 and total (t)Stat3. Bands representing Stat3α, Stat3β, and Stat3δ isoforms are indicated on the right. Heavy lines separate representative samples from individual gels. shows the mean densitometric ratio of (p)Stat3 to (t)Stat3 bands ± standard deviation of the groups (n≥3 for each). Statistically significant (p<0.05) intergroup comparisons are indicated by “*”, “†”, and “§”, respectively.

Article Snippet: The membrane was then stripped (using RestoreTM Western Blot Stripping Buffer, PIERCE, Rockford, IL) and immuoblotting performed to detect total Stat3 protein using mouse IgG monoclonal antibody to Stat3 (BD Biosciences, Rockville, MD; 1∶1000 dilution) as described before , , .

Techniques: Staining, Western Blot, Negative Control, Positive Control, Standard Deviation