Figure 5.

NAT10 positively regulates the osteogenic differentiation of BMSCs in vitro

(A and B) The ac4C level of total RNA increased in BMSCs infected with lentiviruses overexpressing NAT10 and decreased in BMSCs infected with siNAT10, as determined by dot blot analysis, n = 6. (C) ARS staining showed that overexpressing NAT10 caused more calcium nodule formation (upper panels), and ALP staining increased after overexpressing NAT10 (lower panels) (scale bars: 250 μm), n = 6. (D) NAT10 knockdown by siRNA decreased calcium nodule formation (upper panels) and ALP staining (lower panels) (scale bars: 250 μm), n = 6. (E and G) ARS staining was quantified as the absorbance at 562 nm, n = 6. (F and H) ALP activity was determined as units per gram of protein per 15 min, n = 6. (I) Protein levels of the markers of osteoblast differentiation, RUNX2, and Osterix were increased after overexpressing NAT10 during osteogenic induction (left), and knocking down NAT10 had the opposite effect (right), n = 6. (J and K) Quantification of band intensities, n = 6. All data are presented as the means ± SD. ∗p < 0.05, ∗∗p < 0.01 (n = 3 independent experiments).