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Fig. 1 | Journal of Experimental Orthopaedics

Fig. 1

From: Impact of mechanical stimulation on the chondrogenic processes in human bone marrow aspirates modified to overexpress sox9 via rAAV vectors

Fig. 1

Detection of transgene expression in rAAV-transduced human bone marrow aspirates in static versus dynamic culture conditions. Aspirates (150 μl) were transduced with rAAV-RFP, rAAV-lacZ, rAAV-luc, or rAAV-FLAG-hsox9 (40 μl each vector) or let untransduced for maintenance in dynamic or static culture conditions for up to 28 days (n = 4 independent samples tested in triplicate per vector treatment and culture condition in three independent experiments) as described in the Methods. a Macroscopic views of the aspirates are presented for each cultivation systems (day 21, scale bars: 1.5 cm for the static cultures and 2 cm for the dynamic cultures; insets: day 28), b Detection of RFP expression by live fluorescence (day 21, scale bars: 50 μm; top insets: day 7; bottom insets: day 28), c detection of lacZ expression (day 21) by X-Gal staining (macroscopic views, scale bars: 1.5 cm for the static cultures and 2 cm for the dynamic cultures) and following histological processing of the samples (magnification x20, scale bars: 100 μm, all representative data), d detection of luc expression by analysis of luciferase activity with normalization to total cellular proteins, and e detection of SOX9 (day 28, magnification x40, scale bars: 40 μm; top insets: complete section, magnification x4; bottom insets: H&E staining, magnification x40; all representative data) as described in the Methods. aStatistically significant relative to rAAV-RFP application

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