Fig. 2: Precisely mapping the nucleotide acid in locus 22 and its resident transcriptional factors NFE2:MAF. | Oncogenesis

Fig. 2: Precisely mapping the nucleotide acid in locus 22 and its resident transcriptional factors NFE2:MAF.

From: Identifying a locus in super-enhancer and its resident NFE2L1/MAFG as transcriptional factors that drive PD-L1 expression and immune evasion

Fig. 2: Precisely mapping the nucleotide acid in locus 22 and its resident transcriptional factors NFE2:MAF.

A Cloning and direct sequencing of genetic modified locus 22 by T vectors. B DNA sequences of wild-type and two single cell colonies. One is deletion of TGT and another one is addition of T; C Three single cell colonies (1,2,5) were selected and expanded for experiments. Real-time PCR was performed to analyze the mRNA level of PD-L1 and PD-L2 in sgVector, pooled sg-22 and three single cell colonies separated from pooled sg-22; D Western blot was performed to analyze the protein level of PD-L1 in sgVector, sgC1, pooled sg-22 and three single cell colonies from pooled sg-22; E Immunofluorescence was used to examine the expression and distribution of PD-L1 in sgVector and three single cell colonies from pooled sg-22; F The potential transcription factors at 30 bp DNA region around locus 22 in wild-type and dTGT were analyzed by online programs AnimalTFDB (http://bioinfo.life.hust.edu.cn/AnimalTFDB). All potential transcription factors at each allele were listed. The binding of NFE2:MAF was disrupted after deletion of TGT. Data in (C) and (D) are representative of three independent experiments. Data in (E) are representative of two independent experiments. *p < 0.05; **p < 0.01, ***p < 0.001.

Back to article page