Fig. 2: The role of SNORD49A/B in LNC-SNO49AB processing.

a Schematic representation of C/D box snoRNA (upper) and snoRNP (below). b FBL RIP followed by RT-qPCR analysis of copurified RNA in RS4;11 cells. Representative results are shown from three replicates. c Schematic representation of the tRSA RNA pull-down assay. d Formaldehyde denaturing agarose gel analysis of the in vitro-transcribed tRSA and LNC-SNO49AB. Representative results are shown from three replicates. e The results of western blot showed that FBL, NOP58/56 and 15.5 K proteins in the cell lysis were precipitated by in vitro-transcribed LNC-SNO49AB. f Schematic of deletion mutants of LNC-SNO49AB (upper). The FBL binding capability of truncated LNC-SNO49AB is shown below. Tubulin was used as a negative control. g qRT-PCR analysis of FBL, LNC-SNO49AB, SNORD49A/B and SNHG29 in RS4;11 cells treated with FBL sgRNA. The empty PX458 vector was transfected as control. Data were normalized to GAPDH mRNA. Values are the means ± SEM of three independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001 by Student’s t-test. h Schematic of full-length LNC-SNO49AB (upper) and deletions of box C/D (middle, #1–8, red dashed lines). Northern blot of LNC-SNO49AB and the indicated mutants(bottom), 28S and 18S rRNA were acted as the total RNA controls. Representative results are shown from three replicates. i Schematic of the biogenesis and processing of LNC-SNO49AB.