Aperio cs2 slide scanner system

The cancer tissues (T) and paired adjacent tissues (N) were obtained from Affiliated Hospital of Yangzhou University (Yangzhou University, Yangzhou, China) from 2012 to 2013. None of the patients received any radiochemotherapy before the operation. IHC staining was performed as described previously [37 (link)]. Whole-slide images were captured using an Aperio CS2 slide scanner system (Leica Biosystems, Wetzlar, Germany) at a ×40 magnification. IHC stainings of TM4SF1, integrin α6, and p-FAK were analyzed by using the immunoreactive score (IRS) system. The percentage of positive cells was scored on a scale of 0 to 4: 0 if 0% of tumor cells were positive, 1 if 1–25% of cells were positive, 2 if 26–50% were positive, 3 if 51–75% were positive, and 4 if 76–100% were positive. Staining intensity was scored on a scale of 0 to 3 (3 is the strongest). Final IRS score = (score of the staining intensity) × (score of the percentage of positive cells). Score = 0–3 was considered low expression and score = 4–12 was considered high expression. Furthermore, the study was carried out in accordance with the approved guidelines according to the Ethics Committee at Affiliated Hospital of Yangzhou University (Yangzhou University, Yangzhou).

4 weeks old female BALB/c-nude mice were randomly assigned to different groups. A total of 1 × 10⁶ cells (Eca109-, KYSE-510-, and KYSE-410- related TM4SF1-overexpression or TM4SF1-knockdown/rescue cell lines) were injected via the tail vein of each mice. After 7 days, the mice for the overexpression experiment were further randomly divided into two groups (n = 8 for each group) and administered with or without VS-4718 (50 mg/kg) by oral gavage two times a day. On the last day of the treatment, mice were sacrificed and the lungs were initially examined by the naked eye, and hematoxylin and eosin (HE) staining was performed. Whole-slide images of lung lobes were captured using an Aperio CS2 slide scanner system (Leica Biosystems, Wetzlar, Germany) at a 20× magnification. The metastatic nodules of each slide were counted. All experiments were conducted according to the protocols approved by the Institutional Animal Care and Use Committee.