Supplementary materials for the SSR4 article

Information supplementing the SSR4-CDG article. Supplementary Figure S1. HPTLC analysis of total lipids isolated from red blood cell (RBC) ghosts from SSR4-CDG patients and the healthy control. The plates were stained with orcinol for glycosphingolipid detection. Gb4 (globotetraosylceramide) purified from RBC ghosts. Supplementary Figure S2. Glycosphingolipid composition abundances in the control (grey), Patient 1 (red) and Patient 2 (green), quantified from MALDI-TOF spectra as peak areas relative to the reference (shaded). The nomenclature is explained elsewhere; m/z values are given next to the peak IDs. Supplementary Figure S3. Western blotting analysis of Patient 1 (A) and Patient 2 (B) RBC ghosts using anti-glycophorin A antibodies (clones: N61, 6A7, Pep80). N61 recognizes glycophorin A carrying N blood group determinant of MNS system (ISBT No. 002) and glycophorin B. 6A7 binds glycophorin A carrying M blood group determinant of MNS system. Pep80 recognizes glycophorin A irrespective of M/N determinants. GPA, glycophorin A; GPB, glycophorin B. Supplementary Figure S4. Full results of linkage specific sialic acid-derivatizated N-glycomics, with oligomannosidic glycans omitted. Values are relative to the same reference as Fig. 1 (sum of the relative areas of the three Hex2Gn2NeuAc2 glycans). Supplementary Table S1. Sequences of primers used for genotyping. Supplementary Table S2. PCR conditions used for amplification of the A4GALT fragment encompassing the SNP (rs5751348) described by Lai et al. (2014). Supplementary Materials and Methods. Supplementary Materials and Methods.本材料为SSR4-CDG研究论文的补充资料。 补充图S1：针对SSR4-CDG患者与健康对照的红细胞影（red blood cell ghost, RBC ghost）中提取的总脂质，采用高效薄层色谱（high-performance thin-layer chromatography, HPTLC）进行分析，以苔黑酚（orcinol）染色实现糖鞘脂的检测。Gb4（globotetraosylceramide，球四糖基神经酰胺）由红细胞影纯化获得。 补充图S2：以相对于参考物的峰面积为量化指标（阴影部分），通过基质辅助激光解吸电离飞行时间质谱（matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, MALDI-TOF MS）对对照（灰色组）、患者1（红色组）与患者2（绿色组）的糖鞘脂组成丰度进行定量。相关命名规则另有说明；峰标识旁标注了质荷比（m/z）数值。 补充图S3：采用抗血型糖蛋白A（glycophorin A, GPA）抗体（克隆号：N61、6A7、Pep80）对患者1（A子图）与患者2（B子图）的红细胞影进行蛋白质印迹（Western blotting）分析。其中，N61可识别携带MNS血型系统（ISBT编号：002）N血型抗原表位的血型糖蛋白A以及血型糖蛋白B（glycophorin B, GPB）；6A7可结合携带MNS血型系统M血型抗原表位的血型糖蛋白A；Pep80可识别所有血型糖蛋白A，不受M/N抗原表位限制。 补充图S4：展示了连接键特异性唾液酸衍生化N-糖组学的完整结果，省略了寡甘露糖型聚糖。所有数值均以与图1相同的参考物为基准，即三种Hex2Gn2NeuAc2聚糖的相对面积总和。 补充表S1：用于基因分型的引物序列。 补充表S2：用于扩增包含Lai等人（2014年）描述的单核苷酸多态性（single nucleotide polymorphism, SNP）rs5751348的A4GALT基因片段的聚合酶链式反应（polymerase chain reaction, PCR）反应条件。 补充材料与方法。补充材料与方法。