miR-210 and myogenic differentiation 1

miR-210 and myogenic differentiation Table S1 PCR primers for mRNA quantification Gapdh FWD Gapdh REV Rpl13 FWD Rpl13 REV Myod FWD Myod REV Myog FWD Myog REV Myh2 FWD Myh2 REV Des FWD Des REV

5’- TGCCAAGTATGATGACATCAAGAAG - 3’ 5’- GGTCCTCAGTGTAGCCCAAGAT - 3’ 5’- CTCGGCCGTTCTCCTGTAT - 3’ 5’- GTGGAAGTGGGGCTTCAGTA - 3’ 5’- ATGCTGTCTTTTCCCCAAA - 3’ 5’- AACAAGCATGCCAGGATACA - 3’ 5’- CCTTGCTCAGCTCCCTCA - 3’ 5’- TGGGAGTTGCATTCACTGG - 3’ 5’- AGAGCAAAGATGCAGGGAAA - 3’ 5’- TAAGGGTTGACGGTGACACA - 3’ 5’- GCCACCTACCGGAAGCTACT - 3’ 5’- GCAGAGAAGGTCTGGATAGGAA - 3’

Table S2 PCR primers for ChIP assay

miR-210 promoter fragment containing HRE3 and 4 FWD 5’- CACAGCCAGATGGACACACATC - 3’ REV 5’- GCTCCCCTTCGGGAACTCATA – 5’ miR-210 promoter fragment containing HRE1 and 2 FWD 5’- CAGGGAGACACTGTCCAGCG -3’ REV 5’- CTGAAGGGCTGCAGCCAGTG -3’ Table S3 PCR primers for miR-210 promoter luciferase plasmids generation Universal 5’- CCCAAGCTTGGGCTCATCTCCTCTAACTTG - 3’ REV Construct A 5’- GAAGATCTTCAGAGAAGTGAGTGAGCTGGG - 3 FWD Construct B 5’- GAAGATCTTCAGGATCTCCAGGCAAGTA - 3’ FWD Construct C 5’- GAAGATCTTCATGGTGTCCCTGGCCT - 3’ FWD Construct D 5’- GAAGATCTTC CAGGGAGACACTGTCCAG - 3’ FWD Construct E 5’- GAAGATCTTCTCGCCCGCGAAGGGTTGC - 3’ FWD Construct F 5’- GAAGATCTTCCCGCAGCTGAAGTTGGGC - 3’ FWD Construct 5’- GAAGATCTTCAGCCCCGCCCAGTGGGTGGCGGCCCCT - 3’ Myod mut1 Construct 5’- GAAGATCTTCAGCCCCGCCCAGCGGGAGGCGGCCCCT - 3’ Myod mut2 Mutated sequences are underlined

1

miR-210 and myogenic differentiation

Table S4 PCR primers for miR-210 promoter methylation study FWD 1 REV 1 FWD 2 REV 2 FWD 3 REV 3

5’ – GAGGAAAAGGTATGTTTTGGGTTGTA – 3’ 5’ - CCCCCACCCTCATCTCCTCTAACTT – 3’ 5’ - GAGGAAAAGGTATGTTTTGGGTTGTA – 3’ 5’ -CCCCATATACAAATAATCTATCTT – 3’ 5’ - AAGATAGATTATTTGTATATGGGG – 3’ 5’ - CCCCCACCCTCATCTCCTCTAACTT – 3’

2

miR-210 and myogenic differentiation

SUPPLEMENTARY FIGURE LEGENDS Figure S1. miR-210 expression is induced upon rat and quail myoblast differentiation. L6C5 rat myoblasts were cultured in GM and then switched to DM for the indicated time. Then, RNA was extracted and miR-210 (A) and miR-1 (B) levels were measured by qPCR (n=4; # p< 0.02; ** p