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Fig. 5 | Stem Cell Research & Therapy

Fig. 5

From: ER stress arm XBP1s plays a pivotal role in proteasome inhibition-induced bone formation

Fig. 5

Forced expression of XBP1s induces osteoblast differentiation. mMSCs and MC3T3-E1 cells transduced with lentiviral-XBP1s vectors were cultured in the absence or presence of 2.5 μg/mL of doxycycline to induce a higher level of XBP1s expression. Cells transduced with empty lentiviral vectors were used as control. a Alizarin red staining of the mineralized nodule in mMSCs and MC3T3-E1 cells overexpressing XBP1s. The transduced mMSCs and MC3T3-E1 cells were treated with or without doxycycline (Dox) for 5 days and then determined by Alizarin red staining. Images were viewed by a Nikon Eclipse TS100 microscope (scale bars represent 50 μM). b Alkaline phosphatase staining of the transduced mMSCs and MC3T3-E1 cells treated with or without doxycycline for 5 days. c Western blotting analysis of the osteogenic markers in mMSCs and MC3T3-E1 cells overexpressing XBP1s. mMSCs or MC3T3-E1 cells transduced with lentiviral-XBP1s vectors were cultured in the absence or presence of doxycycline for 24 h. d Western blotting analysis of the expression of XBP1s, Atf4, and ER stress markers in the transduced MSCs and MC3T3-E1 cells treated by doxycycline for 24 h. Each assay represents a separate experiment carried out in triplicate

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