Fig. 3.

Effects of circPICALM in BC in vitro and in vivo.

a. Schematic illustration of two siRNAs targeting the junction site of circPICALM. b. and c. Expression of circPICALM and PICALM mRNA in T24 and UM-UC-3 cells transfected with si-NC or si-circPICALM. d. Cell migratory and invasive abilities of BC cells treated with circPICALM siRNAs were measured by migration and invasion assays. Scale bar, 100 μm. e. A representative image of in vivo footpad model. f. BALB/c nude mice were injected with circPICALM overexpression or control UM-UC-3 cells into the footpads. Four weeks later, the popliteal LNs were harvested and measured. (n = 5) g. HE staining of popliteal LNs. Scale bar, 200 μm for the left, 50 μm for the right. h. Distribution of IARs within PICALM gene locus. Upper band showed the whole length of PICALM, while the lower band showed the ±500 bp of exon 9 to exon 12 region. Alignment scores were indicated with specific colours. i. Expression levels of DHX9 were evaluated by western blotting after transfected with DHX9 siRNAs in T24 and UM-UC-3 cells. i. Relative levels of circPICALM in T24 and UM-UC-3 cells were measured after DHX9 was silenced. (Data are presented as the mean ± SD, n = 3. Unpaired, two-tailed student's t-test, **P<0.01).