Fig. 3

SNORA56 promotes 28 S rRNA maturation and translation in CRC cells. (A) Schematic representation of the interaction between SNORA56 and 28 S rRNA, and the mutated regions. (B) PyMOL analysis of the ribosomal structure and U1664, the 28 S rRNA site that is pseudouridylated by SNORA56. (C) A schematic diagram of the primers used to detect total and precursors of 18 and 28 S rRNA. Paired primers were displayed in the same color. (D-E) RT-qPCR analysis was used to reveal the relative levels of unprocessed 28 S rRNA and SNORA56 in HT29 sgNC and sgSNORA56 cells transfected with SNORA56-WT, MUT1, or MUT2. (F) Polysome profiling of HT29 cells with stable SNORA56 knockdown vs. the control. (G) OP-puro analysis of HCT29 cells with stable SNORA56 knockdown vs. the control. (H–K) CCK8 and colony formation assays of the treated HT29 and HCT8 cells. (L) Tumors from nude mice subcutaneous xenografts bearing HT29 sgNC, sgSNORA56, together with empty vector (EV), SNORA56-WT, MUT1 and MUT2, respectively. (M–N) Tumor weights and growth curves. (O) IHC analyses of Ki67 in the xenograft tumors. Scare bar: 100 μm