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. 2020 Jan 19;53(2):e12738. doi: 10.1111/cpr.12738

Figure 4.

Figure 4

YY1 transcriptionally upregulated SPAG5‐AS1/SPAG5 in HG‐treated HPCs. A‐B, 20 transcription factors (TFs) potentially binding to the promoter of SPAG5‐AS1/SPAG5 according to the prediction of human TFDB and PROMO were presented by Venn diagram and analysed via pull‐down analysis under NG, MA or HG treatment. C, Western blot analysis of the YY1 level under NG, MA or HG treatment in cytoplasmic or nuclear fraction of HPCs. D, 3 YY1 binding sites on the promoter of SPAG5‐AS1/SPAG5 were predicted by JASPAR tool. E, ChIP analysis for the DNA fragments with binding site 1, 2 or 3 in the precipitated products of YY1. F, YY1 protein was detected by Western blot in the pulldown of WT SPAG5‐AS1/SPAG5 promoter and SPAG5‐AS1/SPAG5 promoter with site 1, 2 or 3 in HG‐treated HPCs. G, Luciferase reporter analysis elucidated the influence of the YY1 silence on the luciferase activity of SPAG5‐AS1/SPAG5 promoter reporter of each group. H, RT‐qPCR result for the levels of YY1 mRNA, SPAG5‐AS1 and SPAG5 mRNA responding to knockdown of YY1 in HG‐treated HPCs. I, Western blot of YY1, SPAG5, p‐AKT (tthr308 and ser2448), t‐AKT, p‐mTOR and t‐mTOR of each group. All experiments were conducted in triplicates. Data are presented as mean ± SD. *P < .05, **P < .01