Fig. 6. Cell proliferation was retarded when levels of GC UNC-45 mRNA were reduced. (A) Treatment of C2C12 cultures with 2.5 µM GC UNC-45 antisense oligonucleotides suppressed GC UNC-45 mRNA expression by 50%. Northern blot of GC UNC-45 mRNA in 10 µg of total RNA from proliferating cells treated 3 days with none (0), a negative control (C), GC UNC-45 antisense (G) or SM UNC-45 (S) antisense oligonucleotide. Note that SM UNC-45 mRNA is not expressed in proliferating cells (see Fig. 5). GAPDH labeling demonstrated equal RNA loads per lane. (B) Phase-contrast images of proliferating C2C12 cells treated with a negative control or GC UNC-45 antisense oligonucleotide, demonstrating reduced proliferation in GC UNC-45 antisense treated cultures. Cells treated with no or SM UNC-45 antisense oligonucleotide resembled control cultures. Bar, 50 µm. (C) The relative reduction in cell proliferation caused by 3 days of GC or SM UNC-45 antisense treatment, determined from pair-wise comparisons with cultures treated with the control oligonucleotide. Total DNA content was used to gauge cell number. The mean of five experiments is given ±s.d.

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