Figure 1

ULBP3 expression in tumor cell lines and tumor tissues.

(A) Surface expression of ULBP3 on CD133⁻SW620, CD133⁺SW620, K562, 7721, A549 and ECA109 cell lines was measured by flow cytometry (FCM) (n = 3). ULBP3 protein was detected with the monoclonal antibody B2-F1-F1 (5 μg/ml) and a FITC-conjugated goat anti-mouse antibody. On each histogram, the red regions represent the isotype controls and the black lines represent the marker of interest. (B) ULBP3 expression in single-cell suspensions of fresh tumor tissues from colorectal cancer (n = 3), liver cancer (n = 3), lung cancer (n = 3) and gastric cancer (n = 3) and paired adjacent non-tumor tissues (ANTT) was detected by FCM. Representative dot graphs are shown. The results are expressed as the mean ± SEM. (C) Representative results from immunohistochemical staining for ULBP3 in colon, liver, lung and stomach tumor tissues and ANTT are shown. Brown regions represent positive expression of ULBP3. Control indicates ANTT. The slides were analyzed under a 10 × 40 microscope equipped with a camera. (D) Total mRNA was extracted from tumor cell lines, colorectal tumor tissues and ANTT using TRIzol reagent. The mRNA expression of ULBP3 and β-actin was evaluated using RT-PCR. PCR products were visualized by gel electrophoresis in the presence of ethidium bromide. The data are representative of results obtained from at least 3 independent experiments.