Fig. 4

Alizarin red staining of the direct co-culture of intact primary OB and transduced EC after 14 days of induced osteogenic differentiation. (A) Illustration of the Notch signaling pathway. The components of the Notch signaling pathway that were inhibited by lentiviral constructs are highlighted in red, while the activated components are shown in green. (B-C) The results of alizarin red staining after direct co-cultivation of OB with modified EC in the OM. The EC were previously modified 48 h prior to co-cultivation with OB using inhibitory (B) and activating (C) lentiviral constructs targeting components of the Notch signaling pathway. As a control of the lentiviral transduction, the shGFP vector was used, the target gene of which is not expressed in cells, as well as cells without viral load. (D) Quantitative assessment of the intensity of alizarin red staining by spectrophotometry. The results were analyzed using ANOVA, *p < 0.05, **p < 0.01, ****p < 0.001 and compared them relative to the control co-culture there EC were transduced by shGFP, the target gene of which is not expressed in the EC (yellow column)