1. Supplementary Figure S1 Cell index 1224364860728496 hours 1 2 3 4 5 6 Cell index 1224364860728496 hours 1 2 3 4 MiaPaca-2 cells vehicle ABTL0812 A549 cells ABTL0812 Supplementary Figure S1. ABTL0812 inhibits cell proliferation. A549 and MiaPaca-2 cells, seeded in L8 E-Plates, were treated with either vehicle (blue circles and line) or 50  M ABTL0812 (red circles and line). Impedance, which correlates with plate surface covered by attached cells, was monitored in real time during 96 h on a xCELLigence System (Acea Biosciences). Electronic readout of cell-sensor-detected impedance is displayed as arbitrary units called Cell Index. Arrows denote the time when vehicle or ABTL0812 were added to the cells. Each value is the mean ± SD of three different determinations performed in parallel experiments.

2. FL3 Log (PI fluorescence) Counts Vehicle ABTL0812 A549 cells MiaPaca-2 cells 600 400 200 0 400 200 0 1010 2 10 3 1 1010 2 10 3 1 Counts 40 20 0 1010 2 10 3 1 1010 2 10 3 1 50 25 0 Counts Vehicle ABTL0812 FL3 Log (PI fluorescence) Supplementary Figure S2 Supplementary Figure S2. ABTL0812 induces cell death. A549 and MiaPaca-2 cells were treated with either vehicle or 100  M ABTL0812 for 24 h. Cells were collected and incubated with 1  g/ml propidium iodide (PI), and analyzed using flow cytometry. Similar results were obtained in three separate experiments. Lower histograms show percentage of death cells (PI positive). ***p<0.001 from vehicle-treated cells A549 cells MiaPaca-2 cells 20 40 60 80 0 % cell death ( Propidium iodide-positive cells) vehicle ABTL0812 ***

3. 200 ABTL-0812 400600 Vehicle A MiaPaca-2 A549 B 200400600 100 STP0 cleaved Caspase 3 15 Actin PARP-1 cleaved PARP-1 A549 cellsMiaPaca-2 cells DEVDase activity (A.U.F/µg protein) 100 200 300 400 25 50 75 100 125 200400600STP0 STP 1µM Supplementary Figure S3 Supplementary Figure S3. ABTL0812 does not induce apoptotic cell death. (A) Cells were treated with the indicated doses of ABTL0812 or 1µM staurosporine (STP, as a positive control for apoptosis) for 24 h. Cells were fixed and nuclei stained with Hoechst. Cell nuclei were visualized with a Nikon ECLIPSE TE2000-E microscope equipped with epifluorescence optics. Representative pictures showing normal nuclear morphology of ABTL0812-treated cells, and apoptotic nuclear morphology (condensed or fragmented nuclei) in STP-treated cells. (B) Cells treated with the indicated amounts of ABTL0812 (  M) or 1  M STP for 24 h were lysed, and Caspase-3-like activity was measured using the fluorogenic Ac-DEVD-AFC reagent (histograms) in a Bio-Tek FL 600 Fluorometer. Caspase-3 activity was calculated as the increase in arbitrary fluorescence units per microgram of protein. Each value is the mean ± SD of three different determinations, each performed in triplicate, and normalized per microgram of protein. Cells were also analyzed for capase-3 activation by monitoring levels of the cleaved caspase-3 and PPAR-1 by immunoblotting (lower panels). Actin was used as loading control.

4. MiaPaCa-2 A549 VehicleABTL0812 Supplementary Figure S3. ABTL0812 induces LC3 association with autophagosomes. Cells were treated with vehicle (ethanol) or 50  M ABTL0812 for 12 h, before immunostaining for endogenous LC3 (green) and nuclei (Hoechst, blue). Punctua represent autophagosome formation. Supplementary Figure S4

5. PPARα PPAR  0.1110100 1000 25 50 75 100 Concentration ABTL0812 (  ) IC50: 6.8 µM Ki: 7.1 µM % Radioligand binding 0.11101001000 25 50 75 100 Concentration ABTL0812 (  ) IC50: 2.5 µM Ki: 4.7 µM % Radioligand binding Supplementary Figure S5 Supplementary Figure S5. ABTL0812 binds PPARα and PPAR  ligand binding domains Purified human recombinant PPARα or PPAR  were incubated with ABTL0812 and labeled PPAR  agonist 3 H-WY14643 (200 nM) or PPAR  agonist 3 H-Rosiglitazone (5nM). Values are the mean ± SD of two separate determinations performed in duplicate. Ki values were calculated using the Cheng-Prussoff equation (7).

6. A549 Cells MiaPaca-2 cells Supplementary Figure S6 PPRE-Lucierase activity (fold induction) 1 3 5 7 vehicleWY-1463 Rosiglitazone ABTL0812 vehicleWY-1463 Rosiglitazone ABTL0812 1 3 5 7 PPRE-Lucierase activity (fold induction) *** Supplementary Figure S6. ABTL0812 induces PPRA  and PPAR  transcriptional activities. Luciferase-PPAR response element (PPRE) reporter and pRL-CMV-Renilla plasmids were transiently transfected in A549 and MiaPaca-2 cells. 16 h post-transfection, cells were treated with vehicle, PPAR  agonist WY-14643 (1  M) or PPAR  agonist rosiglitazone (1  M) or 50µM ABTL0812. After 24 h, lysates were subjected to a dual-luciferase reporter assay. Each value is the mean ± SD of three different experiments, each performed in triplicate, and normalized using the Renilla values. ***P<0.001 from cells treated with vehicle.

7. 0.0 0.5 1.0 1.5 2.0 2.5 TRIB3 mRNA levels (fold from vehicle of sample 1) Vehicle ABTL0812 Sample 1 Sample 2Sample 3 Sample 4 0.0 0.5 1.0 1.5 2.0 ** GAPDH TRIB3 S1 S2 - + VehicleABTL - + Supplementary Figure S7 Supplementary Figure S7. ABTL0812 induces TRIB3 mRNA levels in Peripheral blood mononuclear cells (PBMC). PBMCs purified from four different human samples were treated for 6 h with vehicle (white columns) or 100  M ABTL0812 (blue columns). After purifying total RNAs with Total RNA Mini Kit Blood/Cultured Cell (Geneaid, New Taipei City, Taiwan), TRIB3 mRNA levels were monitored by RT-qPCR. Value of sample 1 was arbitrarily set to 1. Inset upper panel: data correspond to the mean ± SD of samples 1 to 4, and it shows the mean fold change relative to non-treated cells; (n=4;**P < 0.01 from non-treated cells). Inset lower panel: TRIB3 mRNA levels (as determined by RT-PCR) from ABTL-0812-treated (+) or non-treated (-) samples 1 and 2. ABTL TRIB3 mRNA levels (fold from vehicle)