FIG. 2.
Expression of muscle and urothelial lineage-specific transcripts in differentiated human BMSCs by reverse transcriptase–polymerase chain reaction on day 14. (A) Smooth muscle-specific primers (α-SMA, desmin, calponin, MHC) used on RNA extracted from human BMSCs (p4) induced with human bladder SMC-derived conditional medium for 14 days. Lane 1, fresh smooth muscle tissue; lane 2, cultured SMCs as positive controls; lane 3, cultured urothelium as negative control; lane 4, noninduced BMSCs as control; lane 5, induced BMSCs by coculture with SMCs; lane 6, induced BMSCs by SMC-CM; lane 7, no template control. (B) Urothelial-specific primers (Up-Ia, CK-7/13) used on RNA samples from human BMSCs (p4) induced by coculture with urothelium for 7 days. Lane 1, cultured urothelium as positive; lane 2, noninduced BMSCs as control; lane 3, BMSCs cocultured with urothelium; lane 4, BMSCs cultured using urothelium-CM; lane 5, no template (H2O) control. GAPDH was used as the housekeeping gene for load control. Note: Threefold excess of the reaction mixture was loaded for all the primers in (A) compared with that in (B). Moreover, the bands for all the primers in (A) are overexposed, whereas the bands in (B) are under exposured. α-SMA, alpha smooth muscle actin; MHC, myosin heavy chain; Up-Ia, uroplakin-Ia; CK-7/13, cytokeratin-7/13; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.