#	Aladdin ID	Assay Type	Description	Organism	Compounds	Reference	BAO Format	Source
1.	ALA1169227	B	Inhibition of ERN1 at 1 uM		6	ALA1165965	single protein format	Scientific Literature
2.	ALA1174417	B	Inhibition of ERN1 at 10 uM		2	ALA1177794	single protein format	Scientific Literature
3.	ALA1244834	B	Binding affinity to ERN1		7	ALA1240346	single protein format	Scientific Literature
4.	ALA1827279	B	Activity of ERN1 kinase at 1 uM		1	ALA1821651	single protein format	Scientific Literature
5.	ALA1837987	B	Inhibition of human IRE1 in HL60 cells lysate at 10 uM using post probe-labeling by LC-MS/MS analysis relative to control	Homo sapiens	5	ALA1833925	cell-based format	Scientific Literature
6.	ALA1908532	B	Binding constant for ERN1 kinase domain		72	ALA1908390	single protein format	Scientific Literature
7.	ALA1942750	B	Inhibition of human IRE1 in HL-60 cells lysate assessed as reduction of labeling of acyl-phosphate ATP probe at 100 nM	Homo sapiens	4	ALA1938230	cell-based format	Scientific Literature
8.	ALA2213994	B	Inhibition of ERN1 assessed as residual activity at 10 uM relative to control		1	ALA2203081	single protein format	Scientific Literature
9.	ALA2344390	B	Inhibition of ERN1 (unknown origin)	Homo sapiens	1	ALA2331331	single protein format	Scientific Literature
10.	ALA3111966	B	Inhibition of ERN1 (unknown origin) assessed as residual activity at 10 uM after 1 hr by qPCR analysis relative to control	Homo sapiens	1	ALA3108735	single protein format	Scientific Literature
11.	ALA3265635	B	Inhibition of human recombinant puritin-His-tagged IRE-1 RNase expressed in SF21 cells using XBP-1 RNA stem loop as substrate incubated for 30 mins prior to substrate addition measured after 2 hrs by FRET-suppression assay	Homo sapiens	39	ALA3259679	cell-based format	Scientific Literature
12.	ALA3270871	B	Inhibition of ERN1 (unknown origin) at 1 uM relative to control	Homo sapiens	3	ALA3259739	single protein format	Scientific Literature
13.	ALA3375435	B	Inhibition of human IRE1alpha using 5'[6FAM]-GAGUCCGCAGCACUC-[BHQ1]3' substrate by biochemical fluorescence quenching assay	Homo sapiens	19	ALA3352477	single protein format	Scientific Literature
14.	ALA3412168	B	Inhibition of IRE-1 (unknown origin)	Homo sapiens	3	ALA3407336	single protein format	Scientific Literature
15.	ALA3624354	B	Inhibition of human recombinant IRE1	Homo sapiens	1	ALA3621115	single protein format	Scientific Literature
16.	ALA3705844	B	In Vitro Enzyme Assays: IRE-1 alpha, T1 RNase, and RNase A assays carried out in vitro with several o-vanillin derivatives to demonstrate selectivity of the derivatives for IRE-1 alpha. T1 RNase was assayed as follows. Five ul of a reaction mixture comprising 1x reaction buffer (5x reaction buffer is 100 mM Hepes pH 7.5, 250 mM KOAc, 2.5 mM MgCl2), 3 mM DTT, and 0.4% polyethylene glycol water were added to each well of 384 well plates. Twenty-five nanoliters of a 1 mM test compound solution were added to test wells. Three ul of a 1/48,000 dilution of an approximately 200,000 U/ml RNase T1 (Worthington) preparation were added to each test well and to positive control wells (final concentration 49.5 pg/well). Negative control wells contained only reaction mixture and test compound. After spinning the plates at 1200 rpm for 30 seconds, 3 ul of the mini-XBP-1 mRNA stem-loop substrate described in Example 1 were added to each well of a control plate.	Homo sapiens	271	ALA3638709	single protein format	BindingDB Database
17.	ALA3705669	B	Inhibition Assay: A fusion protein comprising glutathione S transferase (GST) and human IRE-1alpha (GST-IRE-1alpha) obtained from a 500 ml baculovirus-infected insect cell culture can be used to measure IRE-1alpha activity in vitro. Five ul of a reaction mixture comprising IX reaction buffer (5x reaction buffer is 100 mM Hepes pH 7.5, 250 mM KOAc, 2.5 mM MgCl2), 3 mM DTT, and 0.4% polyethylene glycol water is added to each well of 384 well plates. Twenty-five nanoliters of a 1 mM test compound solution are added to test wells. Three ul of a 128 ng/ml IRE-1alpha preparation are added to each test well and to positive control wells (final concentration 5.82 ng/well). Negative control wells contain only reaction mixture and test compound.After spinning the plates at 1200 rpm for 30 seconds, 3 ul of an IRE-1alpha human mini-XBP-1 mRNA stem-loop substrate 5'-CAGUCCGCAGCACUG-3' (SEQ ID NO:1), labeled with the fluorescent dye Cy5 at the 5' end and Black Hole Quencher 2.	Homo sapiens	137	ALA3639376	single protein format	BindingDB Database
18.	ALA3802260	B	Inhibition of partial length wild type human ERN1 expressed in mammalian system assessed as enzyme activity at 1 uM relative to control	Homo sapiens	1	ALA3797029	assay format	Scientific Literature
19.	ALA3887866	B	In Vitro Enzyme Assay: A fusion protein comprising glutathione S transferase (GST) and human IRE-1α (GST-IRE-1α) was obtained from a 500 ml baculovirus-infected insect cell culture and used to measure IRE-1α activity in vitro. Five μl of a reaction mixture comprising 1× reaction buffer (5× reaction buffer is 100 mM Hepes pH 7.5, 250 mM KOAc, 2.5 mM MgCl2), 3 mM DTT, and 0.4% polyethylene glycol water were added to each well of 384 well plates. Twenty-five nanoliters of a 1 mM test compound solution were added to test wells. Three μl of a 128 ng/ml IRE-1α preparation were added to each test well and to positive control wells (final concentration 5.82 ng/well). Negative control wells contained only reaction mixture and test compound.After spinning the plates at 1200 rpm for 30 seconds, 3 μl of an IRE-1α human mini-XBP-1 mRNA stem-loop substrate 5′-CAGUCCGCAGCACUG-3′ (SEQ ID NO:1), labeled with the fluorescent dye Cy5 at the 5′ end and Black Hole Quench		277	ALA3886481	single protein format	BindingDB Database
20.	ALA3887867	B	Cell-Based Assay: Initial cell-based XBP-1 mRNA splicing assays confirmed IRE-1α inhibition with several potent 5-bromo and 6 bromo o-vanillins. HEK293 cells were incubated with compound either overnight or for 2 hours prior to IRE-1α activation with the UPR inducing reagent thapsigargin. IRE-1α mediated XBP-1 splicing was measured by RT-PCR using XBP-1 specific primers flanking the 26 bp intron excised by IRE-1α. The results are shown in FIG. 1. It can be observed that at the higher concentrations, there is relatively more of the unspliced XBP-1 (upper band: substrate) compared to the spliced form (lower band: product).		183	ALA3886481	cell-based format	BindingDB Database