Prognostic Significance of CD200 Protein Expression and its Correlation with COX-2 in Cutaneous Melanoma

Supplementary Figure 1. Expression of CD200 and COX-2 in melanoma tissue. Immunohistochemical staining for (A) CD200 (X 100) and (B) COX-2 (X 40) in melanoma tissue. Paraffin-embedded sections were immunostained with anti-CD200 (R&D systems, Minneapolis, MN, USA; 1:5000) and anti-COX-2 (Neomarkers, CA, USA; 1:50). Double immunofluorescence staining in melanoma tissue of (C) melan-A and CD200 (melan-A, red; CD200, green), (D) melan-A and COX-2 (melan A, red; COX-2, green), and (E) CD200 and COX-2 (CD200, green; COX-2, red). Double-positive cells are indicated by the yellow signals. (F) Correlation between CD200 and COX-2 protein expression in cutaneous melanoma.

Supplementary Figure 2. Progression-free survivals depending on (A) CD200 (B) COX-2 expression in melanoma patients. COX-2, cyclooxygenase-2

Supplementary Figure 3. Effects of COX-2 inhibitor on the expression of CD200 in melanoma cell line. (A, B) Western blotting analysis to evaluate the effect of COX-2 inhibitor on the level of CD200 in Human BRAFV600E A375 melanoma cell. A375 cells were treated with the selective COX-2 inhibitor, celecoxib at different doses. CD200 was downregulated by celecoxib at dose from 60 μM. *p < 0.05, **p < 0.01.

补充图1. 黑色素瘤组织中CD200和COX-2的表达。采用免疫组化染色技术对(A) CD200 (放大倍数：100)和(B) COX-2 (放大倍数：40)在黑色素瘤组织中的表达进行染色。石蜡包埋切片经抗-CD200抗体（R&D systems，明尼阿波利斯，MN，美国；稀释比例：1:5000）和抗-COX-2抗体（Neomarkers，加利福尼亚州，美国；稀释比例：1:50）进行免疫染色。对黑色素瘤组织中(C) melan-A与CD200（melan-A，红色；CD200，绿色）、(D) melan-A与COX-2（melan A，红色；COX-2，绿色）以及(E) CD200与COX-2（CD200，绿色；COX-2，红色）进行双免疫荧光染色。双阳性细胞通过黄色信号标示。(F) 皮肤黑色素瘤中CD200与COX-2蛋白表达的相关性。

补充图2. 根据黑色素瘤患者中(A) CD200和(B) COX-2表达水平，分析无进展生存率。COX-2，环氧化酶-2。

补充图3. COX-2抑制剂对黑色素瘤细胞系中CD200表达的影响。(A, B) 通过Western blotting分析评估COX-2抑制剂对人类BRAFV600E A375黑色素瘤细胞中CD200水平的影响。A375细胞在选择性COX-2抑制剂塞来昔布的不同剂量下进行处理。CD200在60 μM剂量的塞来昔布作用下下调。*p < 0.05，**p < 0.01。