Fig 2. Characterization of circALTO.

(A) The expression of circALTOs and its linear mRNA counterparts after treatment with Actinomycin D at the indicated time points in WaGa cells by qRT–PCR analysis. Both circALTO and linear ALTO were normalized to 18S and then compared to levels at the pre-treatment (0 h) time point. Error bars calculated the SD from 3 biological replicates. The P value was determined using two-tailed t-test. (B) WaGa were fractionated and analyzed by qRT-PCR. MALAT1 and ACTB served as nuclear and cytoplasmic fractionation controls respectively. Values are normalized to the enriched fraction. Error bars were calculated from 3 biological replicates. (C) qRT-PCR of m⁶A or IgG control RNA immunoprecipitation (RIP) of VP-MCC cell lines WaGa. SON, m⁶A RNA IP control. Error bars represent SD (n = 3 biological replicates).