Fig. S2

Fig. S2 Validation of RNLTR12-int expression in MACS isolated OPC (A2B5+) and OL (Mog+), related to Figure 1 and STAR Methods (A) Top: (left) RT-PCR analysis of cytoplasmic beta-actin (Actb) using intron flanking primers. Example shown detection of gDNA contamination in RNA that resulted an upper band. Only single lower band obtained in OPC and OL samples, confirms no gDNA contamination. (right) RT-PCR analysis of RNLTR12-int expression in OPC and OL. PCR product was run on a 2% agarose gel. Bottom: qPCR melting curve analysis resulted only single peak for Actb (left) and RNLTR12-int (right). (B) Actb expressions remained unchanged in OPC and OL as determined by Affymetrix GeneChip. Normalized intensity of all Actb specific probes (17 probes) were used and plotted relative to OPC. Mean ± SEM; n = 9 (OPC), 8 (OL), p > 0.05, Student’s t test (unpaired, two tailed). (C) Expression levels of Mag, Tspan2, and PB1D9 is elevated in OL compared with OPC determined by RT-qPCR. Mean ± SEM; n = 3–4, ∗p < 0.05, ∗∗p < 0.01, Student’s t test (unpaired, two tailed). (D) RNLTR12-int encoded transcript is likely to be a non-coding RNA as predicted by Coding-Non-Coding Identifying Tool (CNIT) algorithm.65 Score of a known protein-coding gene, TATA-box binding protein (Tbp, Transcript ID: ENSRNOT00000002038.4) is shown. (E) RNA-seq reads (GEO: GSM3967160) were aligned to RNLTR12-int consensus sequence, and the depth was plotted.