Vascular SMC Differentiation Marker expression. A and B. Representative immunocytofluorescence of SMC differentiation markers in A10 and A7r5, respectively. Cells were cultured in DMEM supplemented medium for 3 days before immunocytochemistry was performed. Cells stained positive for (a-c) SMA (d-f) SM-MHC, and (g-i) CNN1. The nuclei of were stained with DAPI. Scale bar applies to all images, 250 nM. C. Confocal immunocytochemical staining of SMC differentiation markers in A7r5 cells. Quiesced A7r5 cells and grown in DMEM supplemented with (a-c, g-I, m-o) 5% FBS and (d-f, j-l, p-r) 0.5% FBS for 72 h before cells were analyzed under confocal immunofluorescence microscopy for SMC markers (a-f) SMA and (g-l) SM-MHC and (m-r) CNN1. Arrows designate myofilamentous structures. The nuclei of were stained with DAPI. Scale bar, 100 nm and 250 nm. Data are representative of individual slides with similar results.