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Fig. 1
a, b Results of alkaline phosphatase (ALP) staining and ALP activity assay in Ubap2-knockdown and -overexpressing cells after 5 days with induction of osteoblast differentiation (n = 3 independent experiments; duplicate samples). c, d Results of mineralized nodule formation in Ubap2-knockdown and -overexpressing cells via Alizarin red S (ARS) staining and quantification of ARS-positive cells. Cells were incubated with the osteoblastic medium containing ascorbic acid and β-glycerophosphate for 14 days (n = 3 independent experiments; duplicate samples). e mRNA expression levels of osteoblast differentiation-related genes in Ubap2-knockdown and Ubap2-overexpressing cells. Quantitative reverse-transcription PCR was performed with the specific primers for Runx2, Sp7, and Bglap. Relative quantification of mRNA expression was presented as fold change relative to shCont (Day 5) and normalized to mouse Gusb expression (n = 3 independent experiments; duplicate samples). Control, no induction; Mock, no treatment; shCont, pLKO.1-puro empty vector; and Vector, pDON-5 Neo vector. Statistical differences of multiple groups were determined using one-way analysis of variance (ANOVA), followed by Tukey’s honestly significant difference post hoc test. Exact p-values representing comparison to short hairpin RNA control (shCont) are shown. All data are presented as mean ± SD with individual average values. Source data are provided as a Source Data file.