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. 1998 Mar 23;140(6):1519–1534. doi: 10.1083/jcb.140.6.1519

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Blockade of Ep-CAM–mediated cell–cell interactions causes endocrine differentiation of HFP cells. HFP cells cultured as floating ICCs for 5 d in the presence of either Fab′ fragments of KS1/4 mAb or control Fab′ were used for determination of insulin and glucagon transcript levels in a multiprobe RNase protection assay. A shows that treatment with anti–Ep-CAM KS1/4 Fab′ causes a significant increase of both insulin and glucagon gene transcription. Nicotinamide-treated ICCs (lane NIC) were used in the assay as an internal positive control for induction of endocrine differentiation. Yeast tRNA (lane tRNA) was included as a negative control. Lane UP, undigested probes; M, RNA sizing ladder. Quantitation of band intensities performed by scanning densitometry (B) shows that blockade of Ep-CAM produces a 1.9- and 1.4-fold increase, respectively, of insulin and glucagon mRNAs above the levels measured in control ICCs. Note that the insulin protein content is also increased in ICCs treated with anti–Ep-CAM KS1/4 Fab′ (C). A is representative of four independent assays. Data in B and C are expressed as mean ± SEM of four independent experiments. P values for significant differences were: P < 0.002 for both insulin and glucagon transcript levels compared to control in B; P < 0.005 for samples treated with KS1/4 Fab′ compared to control in C.