Additional file 1: of Geranylgeranylacetone attenuates fibrogenic activity and induces apoptosis in cultured human hepatic stellate cells and reduces liver fibrosis in carbon tetrachloride-treated mice

Regarding the activation of Janus N-terminal kinase (JNK) and p38-mitogen activated protein kinase (MAPK), no significant changes were observed in LX2 cells treated with GGA using western blotting analysis (A). GGA suppressed death receptor 5 (DR5) expression in LX2 cells. The expression of DR5 was assessed by real-time PCR. (B) The upregulation of phosphorylated Akt in LX2 cells treated with GGA was observed using western blotting analysis (C). There was no significant change in the expression of HSP70 in LX2 cells treated with GGA and control cells using western blotting analysis (D). In all experiments, LX2 cells were incubated in the presence or absence of GGA (0.10–0.50 mM) for 24 h. * = P < 0.05. (JPEG 73 kb

Results of RT-PCR and in situ hybridization.

<p>a. Expression level of HBB was significantly higher in carcinoma tissue compare to non-cancerous surrounding tissue. The amount of expression varied highly in cancer tissue. On the other hand, the expression level contributed to relatively narrow range in non-cancerous tissue. b. ISH image from cancer area. HBB mRNA expression (red) was observed in the cancer area. c. ISH fluorescent image from cancer area showed strong positivity for HBB mRNA expression (green).</p

BC-negative case.

<p>a. The color of the epithelia between each dilated IPCL was identical to surrounding mucosa. b. H&E staining histologically confirmed the lesion as intraepithelial neoplasia. c. Immunohistology using anti-Hb Ab showed that the lesion was negative for Hb.</p

Comparison of immunohistological images using anti-Hb Ab.

<p>Immunostaining image showed intense positivity as red blood cells in the cancer area. Non-cancerous mucosa revealed to be negative for anti-Hb Ab.</p

Immunohistological results.

<p>All 36 lesions that were positive for Hb were also positive for Hb immunostaining. Concordance rate of Hb immunopositivity and BC positivity was high as 80.9% (36+2/47).</p

Correlation between BC positivity and the pathology.

<p>97.1% of BC positive lesions were diagnosed as malignant. Overall accuracy of differentiating malignancy from benign pathology was 89.6%.</p

PSP change before and 3 months after POEM.

<p>(a) Conventional white light observation before POEM. Minute whitish stripe is observed on the surface of mucosa. (b) White light with indigocarmine dye. Indigocarmine emphasizes the mucosal pattern. (c) Conventional white light observation 3 months after POEM. Minute stripe was nearly disappeared. (d) Chromoendoscopy image 3 months after POEM. Minute stripe was decreased.</p

Typical findings of primary esophageal achalasia.

<p>(a) Dilation of the esophagus. Dilated esophagus drooped to both sides of the spine. (b) Food remnant in the esophagus. (c) Whitish coating of the mucosa caused by adhesion of the remained food inside of the esophagus and thickening of the mucosa. (d) Functional stenosis of the esophago-gastric junction. Endoscope passes through the tight segment with some resistance. (e) Abnormal contraction of the esophagus. Simultaneous contraction is clearly observed.</p

Comparison between achalasia (left) and non-achalasia (right) patients using indigocarmine and NBI.

<p>Comparison between achalasia (left) and non-achalasia (right) patients using indigocarmine and NBI.</p

Prevalence of the endoscopic findings in patients diagnosed with esophageal achalasia.

<p>EGJ, esophago-gastric junction.</p><p>Prevalence of the endoscopic findings in patients diagnosed with esophageal achalasia.</p