Supplementary Material

Table S1 Listing of the most strongly induced genes during differentiation of human white preadipocytes into adipocytes. A pilot DNA array (Whole Genome, Fa. Agilent) experiment was performed at day 3 of differentiation (see figure 1 for the differentiation scheme) in the presence of ciglitazone. Genes involved in adipocyte function are shown in bold. ACOT2, an example of a rather weakly regulated metabolic gene (involved in fatty acid synthesis), was examined in our experiments and is also included.

Sequence Name / Sequence Description / Accession # / Fold Change
RASD1 / RAS, dexamethasone-induced 1 / NM_016084 / 123.0226
PDK4 / pyruvate dehydrogenase kinase, isozyme 4 / NM_002612 / 98.38912
AOC3 / amine oxidase, copper containing 3 (vascular adhesion protein 1) / NM_003734 / 59.70088
FABP4 / fatty acid binding protein 4, adipocyte / NM_001442 / 58.0028
G0S2 / G0/G1 switch 2 / NM_015714 / 55.8791
ATP1A2 / ATPase, Na+/K+ transporting, alpha 2 (+) polypeptide / NM_000702 / 51.41335
PRKAR2B / protein kinase, cAMP-dependent, regulatory, type II, beta / NM_002736 / 50.4696
APCDD1 / adenomatosis polyposis coli down-regulated 1 / NM_153000 / 41.44944
LPL / lipoprotein lipase / NM_000237 / 41.15984
ITIH5 / inter-alpha (globulin) inhibitor H5 / NM_030569 / 41.10863
THRSP / thyroid hormone responsive (SPOT14 homolog, rat) / NM_003251 / 34.07895
ALDH1L1 / aldehyde dehydrogenase 1 family, member L1 / NM_012190 / 29.85962
PDE8B / phosphodiesterase 8B / NM_003719 / 29.57455
PLEKHA7 / pleckstrin homology domain containing, family A member 7 / NM_175058 / 29.35209
PPP1R1A / protein phosphatase 1, regulatory (inhibitor) subunit 1A / NM_006741 / 23.72021
HES1 / hairy and enhancer of split 1, (Drosophila) / NM_005524 / 22.61243
SCGN / secretagogin, EF-hand calcium binding protein / NM_006998 / 22.36286
GPX3 / glutathione peroxidase 3 (plasma) / NM_002084 / 21.4841
ITIH1 / inter-alpha (globulin) inhibitor H1 / NM_002215 / 20.96656
ADH1A / alcohol dehydrogenase 1A (class I), alpha polypeptide / NM_000667 / 20.61436
ADH1C / alcohol dehydrogenase 1C (class I), gamma polypeptide / NM_000669 / 20.28854
MRAP / melanocortin 2 receptor accessory protein / NM_178817 / 20.2134
SEMA6D / sema domain, transmembrane domain (TM), and cytoplasmic domain, (semaphorin) 6D / NM_153618 / 19.18566
SNIP / SNAP25-interacting protein / NM_025248 / 19.07324
PALMD / palmdelphin / NM_017734 / 18.79423
AGT / angiotensinogen (serpin peptidase inhibitor, clade A, member 8) / NM_000029 / 18.74785
ADIPOQ / adiponectin, C1Q and collagen domain containing / NM_004797 / 18.72321
SLC40A1 / solute carrier family 40 (iron-regulated transporter), member 1 / NM_014585 / 18.37927
MLXIPL / MLX interacting protein-like / NM_032951 / 17.93635
AQP7 / aquaporin 7 / NM_001170 / 17.45023
LIPE / lipase, hormone-sensitive / NM_005357 / 16.86631
TSPAN15 / tetraspanin 15 / NM_012339 / 16.70847
SEMA3G / sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3G / NM_020163 / 16.29909
GPD1 / glycerol-3-phosphate dehydrogenase 1 (soluble) / NM_005276 / 15.8142
CDKN1C / cyclin-dependent kinase inhibitor 1C (p57, Kip2) / NM_000076 / 15.48549
SORT1 / sortilin 1 / NM_002959 / 15.31392
CA3 / carbonic anhydrase III, muscle specific / NM_005181 / 15.24551
ACACB / acetyl-Coenzyme A carboxylase beta / NM_001093 / 15.14941
AQP3 / aquaporin 3 (Gill blood group) / NM_004925 / 15.00957
TRIM63 / tripartite motif-containing 63 / NM_032588 / 14.99371
.... / .... / .... / ....
ACOT2 / Homo sapiens acyl-CoA thioesterase 2 / NM_006821 / 4.61221

Table S2 Listing of the most strongly down-regulated cytokines in the pilot cDNA array experiment described in the legend of Table S1.

Sequence Name / Sequence Description / Accession # / Fold Change
CXCL5 / chemokine (C-X-C motif) ligand 5 / NM_002994 / -13.3
IL8 / interleukin 8 / NM_000584 / -6.1
CXCL6 / chemokine (C-X-C motif) ligand 6 / NM_002993 / -5.6
CCL2 / chemokine (C-C motif) ligand 2 / NM_002982 / -5.2
CCL7 / chemokine (C-C motif) ligand 7 / NM_006273 / -4.8

Fig. S1 Absence of a major effect of metformin on adipocyte differentiation as measured by changes in gene expression. Cells were differentiated as shown in figure 1 in the presence of pioglitazone or metformin; the absence of a test compound served as negative control. The fold changes relative to this negative control are indicated. Metformin-induced changes tended to be the opposites of the pioglitazone effects but did not reach statistical significance (n=4). Data are represented as mean +/- SEM; *, p≤0.05, **, p≤0.01, ***, p≤0.001; two-way ANOVA with Bonferroni post test.


Fig. S2 Effect of the PPARg antagonist T0070907 on the expression of PDK4 and ACOT2, two genes involved in metabolism. Human preadipocytes were differentiated with pioglitazone (10 µM), glimepiride (10 µM) or glibenclamide (2 µM) for six days in the presence or absence of T0070907. The induction of PDK4 and ACOT2 was monitored by quantitative PCR (n=4). Data are given relative to negative control and are represented as mean +/- SEM; *, p≤0.05, **, p≤0.01, ***, p≤0.001; one-way ANOVA with Bonferroni post test.