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. 2022 Dec 15;14(12):8903–8917.

Figure 6.

Figure 6

HOTTIP promotes EFNA3 transcription by recruiting CTCF to the EFNA3 promoter. A: mRNA levels of EFNA3 in mice treated with OVA or/and HOTTIP knockdown were measured by qRT-PCR. B, C: The expression of EFNA3 in mice treated with OVA or/and HOTTIP knockdown was measured by western blot. D: EFNA3 expression in lung tissues of mice treated with OVA or/and HOTTIP knockdown was detected with immunohistochemical staining. E, F: The expression of CTCF and EFNA3 in 16HBE cells with CTCF knockdown were determined by western blot. G, H: The expression of CTCF in 16HBE cells transfected with HOTTIP knockdown were measured by western blot assay. I: RNA pull-down assay was performed to test the binding relationship between HOTTIP and CTCF. J: The interaction between HOTTIP and CTCF was confirmed by RNA binding protein immunoprecipitation (RIP) assay. K: Dual luciferase assay was performed to confirm the interaction between CTCF on site 1 of EFNA3 promoter. L: Dual luciferase assay was performed to confirm the interaction between CTCF on site 2 of EFNA3 promoter. M: The interaction between CTCF and EFNA3 promoter was confirmed by chromatin immunoprecipitation (ChIP) assay. *P < 0.05, **P < 0.01, ***P < 0.001 of one-way ANOVA with Tukey test or Student’s t-test.