Fig. 2
The effect of 810 nm NIR light on osteogenic differentiation was related to the ubiquitination degradation of CRY1. a Immunofluorescence staining of CRY1 (green) in BMSCs with or without 810 nm NIR light irradiation, together with nuclei (blue) (left), and the quantitative analysis of nucleus/cytoplasm fluorescence ratio (right) were represented. b BMSCs were treated with or without 810 nm NIR light for the indicated minutes. The protein expressions of CRY1 and H3 in the nucleus and the protein expressions of CRY1 and GAPDH in the whole cell were analyzed by Western blot. c Immunofluorescence staining of CRY1 (green) in MC3T3-E1 cells with or without 810 nm NIR light irradiation, together with nuclei (blue) (left), and the quantitative analysis of nucleus/cytoplasm fluorescence ratio (right) were represented. d MC3T3-E1 cells were treated with or without 810 nm NIR light for the indicated minutes. The protein expressions of CRY1 and H3 in the nucleus and the protein expressions of CRY1 and GAPDH in the whole cell were analyzed by Western blot. e Representative ALP staining images in scramble BMSCs and Cry1-knockdown BMSCs with or without 810 nm NIR light irradiation for 7 days. f MC3T3-E1 cells were transfected with HA-Ub for 24 h and then treated with or without 810 nm NIR light. The ubiquitination level of CRY1 in the nucleus was detected using an anti-HA antibody. g BMSCs were treated with or without KL001 (1 μg·mL−1) for 2 h before 810 nm light irradiation. The protein expressions of CRY1 and H3 in the nucleus were analyzed by Western blot. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.000 1. Scale bar: 50 μm for (a, c) and 100 μm for (e)
