Journal: Cell Proliferation

Article Title: Circ‐CTNNB1 drives aerobic glycolysis and osteosarcoma progression via m6A modification through interacting with RBM15

doi: 10.1111/cpr.13344

Figure Lengend Snippet: Expression of circ‐CTNNB1 in OS cell lines and tissues, and subcellular location of circ‐CTNNB1. (A) Schematic illustration of the genomic location of circ‐CTNNB1 derived from its host gene and validation by Sanger sequencing. (B) RT‐PCR assay showing the presence of circ‐CTNNB1 with divergent and convergent primers from cDNA or genomic DNA (gDNA) of differential OS cell lines using β‐actin as the negative control. (C) qRT‐PCR analysis of the expression of circ‐CTNNB1 after RNase R treatment in 143B or MG‐63 cells. (D) Northern blot showing the differential expression of circ‐CTNNB1 in hFOB 1.19 cells and OS cell lines. (E, F) qRT‐PCR assay showing the relative levels of circ‐CTNNB1 (normalized to β‐actin) in cultured cell lines and human tissues. (G) RNA‐FISH assay showing the nuclear localization of circ‐CTNNB1. Scale bar: 10 μm. (H, I) TOP/FOP flash assay (H) and Western blot (I) assay revealing the β‐catenin activity and protein levels of CTNNB1 in 143B and MG‐63 cells. (Data were mean ± SEM of three experiments. Student's t ‐test and ANOVA compared the difference in C, E, F, H. * p < 0.05, ** p < 0.01)

Article Snippet: The TOP‐FLASH and FOP‐FLASH reporters for the activity of the canonical Wnt pathway were obtained from Millipore (Temecula, CA, USA).

Techniques: Expressing, Derivative Assay, Sequencing, Reverse Transcription Polymerase Chain Reaction, Negative Control, Quantitative RT-PCR, Northern Blot, Cell Culture, Western Blot, Activity Assay