TABLE 4.

ELISA assays for selected 4-ATQs.^25,26 Primary assays: (a) GLP-1 secretion from mouse secretin tumor (mSTC-1) intestinal enteroendocrine cells (mSTC-1 GLP1 Sec) with percent stimulation determination at 20 μM and 2 μM concentration of test compound. (b) Concentration–response curve (CRC) based determination of the concentration that produces a half-maximum response (EC₅₀) in GLP-1 secretion from mSTC-1 cells (mSTC-1 GLP1 Sec). (c) CRC-based determination of EC₅₀ in GLP-1 release from a human cell line derived from ascites fluid of a colorectal adenocarcinoma (hNCI-H716 GLP1 Sec CRC). (d) Cytolethality counter assay measuring leakage of lactate dehydrogenase (LDH) from mSTC-1 cells (mSTC-1 LDH Sec). (e) Cytolethality counter assay measuring LDH leakage from hNCI cells (Basal hNCI LDH Sec). Secondary assay: GLP-1 secretion in a cell line derived from colonic tumors of transgenic mice expressing large T antigen under the control of the proglucagon promoter (GLUTag GLP1 Sec). ND = not determined.

Entry	Compound	mSTC-1 GLP1 Sec %Stim@20 μM/2 μM	mSTC-1 GLP1 Sec CRC EC50 [μM]	hNCI-H716 GLP1 Sec CRC EC50 [μM]	mSTC-1 LDH Sec EC50 [μM]	Basal hNCI LDH Sec EC50 [μM]	GLUTag GLP1 Sec %Stim@10 μM
1	5a	68/2	5.3	>40	>40	>40	ND
2	5b	98/0	10.8	>40	>40	>40	ND
3	5c	53/0	17.2	>40	>40	>40	ND
4	5d	91/25	1.4	>40	>40	>40	6
5	5e	0	ND	ND	ND	ND	ND
6	5f	22/0	ND	ND	ND	ND	ND
7	6a	40/8	>40	>40	>40	>40	ND
8	6c	0	ND	ND	ND	ND	ND
9	6g	0	ND	ND	ND	ND	ND
10	8d	28/0	>40	>40	>40	>40	ND
11	8e	0	ND	ND	ND	ND	ND
12	8f	0	ND	ND	ND	ND	ND
13	8i	0	ND	ND	ND	ND	ND
14	8j	0	ND	ND	ND	ND	ND
15	8k	53/0	8.3	>40	>40	>40	ND
16	8l	0	ND	ND	ND	ND	ND
17	8m	0	ND	ND	ND	ND	ND
18	8n	49/0	26.1	>40	>40	>40	ND
19	8o	61/0	16.9	>40	>40	>40	ND
20	8p	80/0	21.9	>40	>40	>40	ND
21	8q	18/0	ND	ND	ND	ND	ND
22	9	99/82	2.2	>40	>40	>40	ND