Fig 2: MACF1 promoted bone formation by inhibiting the activities of HES1 in ageing-related osteoporosis. A, Selected area of RNA-seq heat map for femur tissues of 6- and 21-mo male (up) and female (below) C57BL/6 mice. B, Expression of HES1 in femur tissues of ageing-related osteoporosis patients (left) and ageing C57BL/6 mice (right), as detected by RT-PCR (up) and Western blot (below) (mean ± SD, ***P < .001). C, Expression of HES1 in femur tissues of ageing-related osteoporosis patients (up) and ageing C57BL/6 mice (below), as detected by immunohistochemical staining. Scale bar: 50 µm. D, Correlation analysis between Hes1 levels and P16 mRNA levels in femur tissues from C57BL/6 mice, as detected by RT-PCR. E, Expression of HES1 in MACF1 overexpression MC3T3-E1 cells (left) and MACF1 knockdown MC3T3-E1 cells (right), as detected by RT-PCR (up) and Western blot (below) (mean ± SD, ***P < .001). F, Correlation analysis between Macf1 levels and Hes1 mRNA levels in femur tissues from C57BL/6 mice, as detected by RT-PCR. G, Activities of HES1 in MACF1 overexpression MC3T3-E1 cells (left) and MACF1 knockdown MC3T3-E1 cells (right), as detected by luciferase reporter assay (mean ± SD, ***P < .001). H, Rescuing effect of MACF1 overexpression plasmid on HES1 activities of MACF1 knockdown MC3T3-E1 osteoblastic cells, as detected by luciferase reporter assay (mean ±SD, ***P < .001)