Fig. 3
a Alizarin Red staining for calcium nodule formation in the control and Botox-injected group cultured in either basal or osteogenic induction medium (OIM) showing the decreased calcium nodules formation in the Botox group. b Quantitation of Alizarin Red by read at 595 nm of destaining solution by 96-well plate reader and normalised to basal-medium control group (n = 3; mean ± SD; *p < 0.05, **p < 0.005, *** p < 0.001). c mRNA expressions of osteogenic differentiation markers (ALP, BMP2 and RUNX2) in the control and Botox group. Individual mRNA expression levels were first normalised to internal control (36B4) and then normalised against mRNA expression level from the control group. (n = 3; mean ± SD; **p < 0.005, ***p < 0.001)