Fig. 7

Hsa_circ_0003258 promotes PCa metastasis through HDAC4 pathway. A Venn diagram showing the RNA candidates as the targets of IGF2BP3. B. The qRT-PCR showing the mRNA level change of the predicated targets after silencing hsa_circ_0003258. C Western blot showing the protein level of HDAC4 after silencing hsa_circ_0003258. D RIP assays showing the association of IGF2BP3 with HDAC4. Relative enrichment representing RNA level associated with HDAC4 compared to an input control. IgG antibody served as a control. E-F The m6A modification site of HDAC4 predicted by SRAMP website tools and m6A bound to HDAC4 performed by meRIP. G HDAC4 protein expression determined by Western blot. H hsa_circ_0003258 depletion on the stability of HDAC4, cells were treated with Actinomycin D at indicated time points and mRNA levels of HDAC4 were validated with qRT-PCR. The U6 was used as normalization control. I-J QRT-PCR and Western blot assay examined the knockdown efficiency of IGF2BP3 and the protein level of HDAC4 after silencing of IGF2BP3. K IGF2BP3 depletion on the stability of HDAC4, cells were treated with Actinomycin D at indicated time points and mRNA level of HDAC4 were validated by qRT-PCR. The U6 was used as normalization control. L-M Transwell assay and Western blot were used to detect the migratory capacity and the protein level in PCa cells after silencing HDAC4. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001