ERCC4 / XPF内含子的位置变异在日本XP-F情况下确定。(A)估计pre-mRNA产品结构造成ERCC4 / XPF基因内区变体。神秘的内含子片段中确定病人的mRNA蓝线所示。(B) cDNA序列ERCC4 / XPF外显子1 - 2边界在48 br(正常)和XP43NG (XP-F)。5′神秘第1内含子片段所示蓝色的字母。(C) cDNA序列ERCC4 / XPF exon8-intron9边界XP2YO (XP-F)。神秘的基因内区8片段所示蓝色的字母。箭头表示变异的位置。(D)数字定量PCR(数字qPCR)发现减少ERCC4 / XPF表达和基因内区1的产品异常剪接XP43NG(满酒吧、PCR扩增ERCC4 / XPF外显子1 - 2边界;灰色酒吧、PCR扩增5′神秘碎片ERCC4 / XPF基因内区1; open bars, a control PCR product of the TBP gene). (E) Digital qPCR detected the reduction of ERCC4/XPF expression in XP3YO (XP-F) (filled bars, PCR amplifying the ERCC4/XPF exons 1 to 2 boundary; hatched bars, exons 7 to 8 boundary; gray bars, exons 8 to 9 boundary; open bars, TBP). (F) Immunoblotting of the XPF protein. wild type and ΔERCC4/XPF, wild type and ERCC4/XPF-deficient HeLa cells; 48BR, normal; XP24BR, XP-F control; [XP136KO, XP37NG, XP43NG, XP101OS, XP97NG, XP165KO, XP103NG, XP4NG, XP48NG, XP90NG, XP95NG, XP18NG, XP133KO, XP23OS, XP96NG, XP2YOSV40, and XP3YO], the Japanese XP-F cases. b-actin (ACTB) as a loading control. XPF-upper bands represent the stop-loss product, p.*917Rext*83. Credit:美国国家科学院院刊》上(2023)。DOI: 10.1073 / pnas.2217423120
