Fig. 4: LETR1 is a bona fide lncRNA expressing three main transcripts in LECs.
a Cumulative fraction analysis of Coding Potential Assessment Tool (CPAT)54 (left) and Phylogenetic Codon Substitution Frequencies (phyloCSF)55 (right) scores of LETR1 (19 transcripts), GAPDH (21 transcripts, known protein-coding gene), and MALAT-1 (15 transcripts, known lncRNA). P values were calculated using the two-sample Kolmogorov–Smirnov test. b Agarose gel showing the results after 3′ Rapid Amplification of cDNA Ends (RACE) (two replicates). White boxes: two excited bands further processed following the SMARTER 3′ RACE protocol. c Schematic representation of 3′ RACE results depicting the three LETR1 transcripts expressed in LECs: LETR1-1 (~1100 bp), LETR1-2 (~1200 bp), LETR1-3 (~600 bp). RNA-Seq signal was visualized through the Zenbu genome browser123. LETR1 transcript sequences are listed in Supplementary Data 8. d Comparison of qPCR levels of GAPDH (polyA+), H2BK (polyA−), LETR1-1, LETR1-2, LETR1-3 after cDNA synthesis with either oligodT or random hexamers primers in neonatal LECs derived from three donors. e In vitro translation assay results of LETR1-1, LETR1-2, and LETR1-3. A construct containing the luciferase gene (62 kDa) was used as positive control. Uncropped western blot image is shown in Supplementary Figure 9. f Expression levels of LETR1-1, LETR1-2, and LETR1-3 in neonatal LECs derived from three donors. Data are presented as mean values + SD (n = 3 in d and f).
