Fig. 1.

CRISPR/Cas9 deletion of Rorβ in MC3T3-E1 preosteoblastic cells. (A) The intron/exon structure of the mouse Rorβ gene is shown. (B) Exon 2 (in uppercase) of the mouse Rorβ gene was mutated using CRISPR/Cas9 gene editing methodology. The guide RNA (gRNA) sequence used to target the allele is highlighted in yellow. The translated amino acid sequence for each allele of the MC3T3-Cont and MC3T3-ΔRorβ cell lines is shown underneath the respective DNA sequence, illustrating the frameshift mutations in the latter cell line. The dashes in the DNA sequence represent single base pair deletions. (C–G) Both cell models were treated with osteogenic medium at confluence and QPCR for the denoted bone markers genes was performed on samples collected at days 0 to 10 (n = 6). Values are expressed as fold change (mean ± SE) relative to day 0. Values of p are shown for repeated-measures ANOVA.