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. 2023 Jan 15;56(7):e13400. doi: 10.1111/cpr.13400

FIGURE 2.

FIGURE 2

FOXP3 and FUS promote the transcription and maturation of non‐SMC condensin I complex subunit H (NCAPH) respectively. (A) Potential transcription factors for regulating NCAPH expression were predicted by ALGGEN and LASAGNA databases. (B) Luciferase reporter assays were carried out to verify the interaction between FOXP3 protein and NCAPH promoter. ***p < 0.001 versus vector; ### p < 0.001 versus sh‐NC. (C) ChIP assays show the enrichment of FOXP3 protein in NCAPH promoter. ***p < 0.001 versus IgG. (D and E) FOXP3 overexpression or depletion changed the enrichment of FOXP3 protein in NCAPH promoter. ***p < 0.001 versus vector; ### p < 0.001 versus sh‐NC. (F) The effect of FOXP3 on NCAPH protein expression was detected by Western bolting in 786‐O and Caki‐1 cells. (G and H) The effect of FOXP3 on NCAPH mRNA expression was detected by Western bolting in 786‐O and Caki‐1 cells. ***p < 0.001 versus vector; ### p < 0.001 versus sh‐NC. (I) The table listed the hnRNPs which were predicted to bind to NCAPH by the ENCORI database. (J) RIP assay was used to verify the interaction between FUS protein and pre‐NCAPH. ***p < 0.001. (K) Pull‐down assay was further carried out by pre‐NCAPH probes, NCAPH probes, and NC probes to verify the interaction between FUS protein and pre‐NCAPH. (L) The effect of FUS on pre‐NCAPH and NCAPH mRNA expression. ***p < 0.001 versus vector. (M) The effect of FUS on NCAPH protein expression. Each experiment was repeated at least three times.