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Fig. 2 | Cancer Cell International

Fig. 2

From: Long non-coding RNA LINC01116 is activated by EGR1 and facilitates lung adenocarcinoma oncogenicity via targeting miR-744-5p/CDCA4 axis

Fig. 2

EGR1 activates the transcription of LINC01116. A EGR1 expression in LAD cell lines and HBE was investigated by qRT-PCR. One-way ANOVA was applied for analysis. B The expression level of EGR1 in A549 and PC9 cells was quantified subsequent to the transfection with pcDNA3.1/EGR1. Student’s t test was applied for analysis. C The impacts of EGR1 overexpression on LINC01116 was analyzed by qRT-PCR. Student’s t test was applied for analysis. D The DNA binding motif of EGR1 on LINC01116 promoter and three specific binding sites were predicted by JASPAR. E LINC01116 promoter was divided into four sectional parts based on predicted binding sites. F ChIP assay revealed the interaction of EGR1 with P4 sectional area in LINC01116 promoter region. Student’s t test was applied for analysis. G The constructions of full promoter (P FL) and P4 deleted (P D) promoter region were sub-cloned into pGL3 luciferase reporter vector. H Luciferase activity was assessed in HEK-293 T cells co-transfected with pcDNA3.1 or pcDNA3.1/EGR1 and pGL3 luciferase reporter vectors carrying P FL or P D region. Student’s t test was applied for analysis. I The sequence of P4 wild type and corresponding point mutant sequences, including P4-Mut1, P4-Mut2 and P4-Mut3 were sub-cloned into pGL3 reporter vector. J Luciferase reporter assay was conducted for analysis of promoter activity. Student’s t test was applied for analysis. **P  <  0.01

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