Fig. 6: DRG neurons induce cytoplasmic accumulation of β-catenin and its translocation into the nucleus in MSCs | Cell Death & Disease

Fig. 6: DRG neurons induce cytoplasmic accumulation of β-catenin and its translocation into the nucleus in MSCs

From: Dorsal root ganglion neurons regulate the transcriptional and translational programs of osteoblast differentiation in a microfluidic platform

Fig. 6

Sensory neurons derived from rat DRG (5 × 104 cells/cm2) and rat bone marrow MSCs (104 cells/cm2) were cocultured in microfluidic devices for 7 days. DRG neurons were maintained in DMEM supplemented with 2% (v/v) B-27 and 1 μM AraC; MSCs were incubated in OIM composed of DMEM-low glucose with 10% (v/v) FBS, 1 × 10−9 M dexamethasone, 10 mM β-glycerophosphate, and 50 μg/mL ascorbic acid. a Subcellular distribution of β-catenin in MSCs was evaluated at 4 and 7 days of coculture by IF using antibodies directed against β-catenin coupled to Alexa Fluor® 488 (green), and DAPI (nuclei; blue) under a confocal microscope. Scale bar = 100 μm. b–d The level of fluorescence for β-catenin was measured at 4 and 7 days of coculture by using the ImageJ software in each MSC and normalized to the monoculture value on day 4. Data expressed as median, 25 and 75 percentiles, min/max. (n) indicates the total number of cells counted for each group. e Expression profile of Ctnnb1 in MSCs was assessed at 4 and 7 days of coculture by RT-qPCR and depicted as a relative ratio to the housekeeping gene Hprt1 normalized to the monoculture value on day 4. Data expressed as mean ± SD. (n) indicates the total number of samples for each group. *p < 0.05; ***p < 0.001 statistically different from monoculture. The results represent three independent experiments.

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