|  | IL-1β/INF-γ | DNA Content (µg/30 islets) | Insulin Content (µg/µg DNA) | Insulin mRNA Relative Levels |
|---|
CBA control | − | 0.72 ± 0.04 | 900 ± 120 | 1.9 ± 0.4 |
CBA control | + | 0.64 ± 0.08 | 1070 ± 200 | n.d. |
CBA-GTK | − | 0.64 ± 0.08 | 1020 ± 125 | 1.8 ± 0.3 |
CBA-GTK | + | 0.60 ± 0.08 | 1070 ± 100 | n.d. |
- Ten islets in triplets were isolated from control or GTK-transgenic CBA mice of both genders, cultured in RPMI 1640 + 10% serum for 24 hr in the absence or presence of 1000 U/ml INF-γ and 50 U/ml IL-1β and used for insulin release (see Fig. 2). The islets were then pooled, homogenized in 200 µl of water and the insulin and DNA contents were measured. Values are mean ± SEM for 12 observations. Equal amounts of total RNA from 60 islets cultured for 24 hr in RPMI 1640 + 10% serum were run on a denaturing agarose gel and subjected to Northern blotting for insulin and β-actin. The relative insulin mRNA levels were determined by densitometry. Values are mean ± SEM for 10 observations; n.d., not determined.
