Figure S2: Compound induced biomarker expression in HepG2 cells. The presented values (relative expression fold changes) represent mean values that were calculated from 3-5 independent experiments. The error bars illustrate the standard deviation of the independent experiments. A gene was considered to be up regulated when crossing the threshold line, which illustrates a significant increase of at least 2.5 fold change induction.
0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0
1 2 3 4 5 6
APAP induced biomarker expression (n=4) HepG2 cells
Fo ld c h a n ge 2 - Δ Δ C T
0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0
0.5 1 1.5 2 2.5 3
ASP induced biomarker expression (n=3) HepG2 cells
CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32
Concentration [mM]
Fo ld c h a n ge 2 - Δ Δ C T
0 0.51 2.6
12.8 64
30000 0 0.51 2.6
12.8 64
30000 0 0.51 2.6
12.8 64
30000 0 0.51 2.6
12.8 64
30000 0 0.51 2.6
12.8 64
30000 0 0.51 2.6
12.8 64
30000 0 0.51 2.6
12.8 64 30000 0
0.5 1 1.5 2 2.5 3
BPR induced biomarker expression (n=3) HepG2 cells
Fo ld c h a n ge 2 - Δ Δ C T
0 1.6 8 40 200
1000 0
1.6 8 40 200
1000 0
1.6 8 40 200
1000 0
1.6 8 40 200
1000 0
1.6 8 40200
1000 0
1.6 8 40 200
1000 0
1.6 8 40 200 1000 0
2 4 6 8 10 12 14 16 18
CBZ induced biomarker expression (n=3) HepG2 cells
CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32
Concentration [µM]
Fo ld c h a n ge 2 - Δ Δ C T
0 0 0.01
0.06
0.32 90 0 0
0.01 0.06
0.32 90 0 0
0.01 0.06
0.32 90 0 0
0.01 0.06
0.32 90 0 0
0.01 0.06
0.32 90 0 0
0.01 0.06
0.32 90 0 0
0.01 0.06
0.32 90 0
0.5 1 1.5 2 2.5 3 3.5 4 4.5
CHL induced biomarker expression (n=3) HepG2 cells
Fo ld c h a n ge 2 - Δ Δ C T
0 0.06
0.32 1.6 8
400 0
0.06
0.32 1.6 8
400 0
0.06
0.32 1.6 8
400 0
0.06
0.32 1.6 8
400 0
0.06
0.32 1.6 8
400 0
0.06
0.32 1.6 8
400 0
0.06
0.32 1.6 8 400 0
0.5 1 1.5 2 2.5 3 3.5 4 4.5 5
DFN induced biomarker expression (n=3) HepG2 cells
Fo ld c h a n ge 2 - Δ Δ C T
0 0.01
0.06 0.32 1.6
700 0
0.01 0.06
0.32 1.6
700 0
0.01 0.06
0.32 1.6
700 0
0.01 0.06
0.32 1.6
700 0
0.01 0.06
0.32 1.6
700 0
0.01 0.06
0.32 1.6
700 0
0.01 0.06
0.32 1.6 700 0
0.5 1 1.5 2 2.5 3
FAM induced biomarker expression (n=3) HepG2 cells
CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32
Concentration [µM]
Fo ld c h a n ge 2 - Δ Δ C T
0 0.01
0.06
0.32 1.6 40 0 0.01
0.06
0.32 1.6 40 0 0.01
0.06
0.32 1.6 40 0 0.01
0.06
0.32 1.6 40 0 0.01
0.06
0.32 1.6 40 0 0.01
0.06
0.32 1.6 40 0 0.01
0.06
0.32 1.6 40 0
0.5 1 1.5 2 2.5 3 3.5 4 4.5 5
HYZ induced biomarker expression (n=4) HepG2 cells
Fo ld c h a n ge 2 - Δ Δ C T
0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 5
INAH induced biomarker expression (n=4) HepG2 cells
CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32
Concentration [mM]
Fo ld c h a n ge 2 - Δ Δ C T
0 0.06
0.32 1.6 8 40 0 0.06
0.32 1.6 8 40 0 0.06
0.32 1.6 8 40 0 0.06
0.32 1.6 8 40 0 0.06
0.32 1.6 8 40 0 0.06
0.32 1.6 8 40 0 0.06
0.32 1.6 8 40 0
0.5 1 1.5 2 2.5 3
LAB induced biomarker expression (n=3) HepG2 cells
CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32
Concentration [µM]
Fo ld c h a n ge 2 - Δ Δ C T
0
0.32 1.6 8 40200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40200 0
0.32 1.6 8 40 200 0
0.5 1 1.5 2 2.5 3
LEV induced biomarker expression (n=3) HepG2 cells
Fo ld c h a n ge 2 - Δ Δ C T
0 0.02 0.1
0.51 2.6
100 0
0.02 0.1 0.51 2.6
100 0
0.02 0.1 0.51 2.6
100 0
0.02 0.1 0.51 2.6
100 0
0.02 0.1 0.51 2.6
100 0
0.02 0.1 0.51 2.6
100 0
0.02 0.1 0.51 2.6
100 0
0.5 1 1.5 2 2.5 3
MEL induced biomarker expression (n=3) HepG2 cells
CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32
Concentration [nM]
Fo ld c h a n ge 2 - Δ Δ C T
0
0.32 1.6 8 40 200 0
0.32 1.6 8 40200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
5 10 15 20 25 30 35 40 45 50
NFT induced biomarker expression (n=3) HepG2 cells
Fo ld c h a n ge 2 - Δ Δ C T
0
0.32 1.6 8 40 330 0
0.32 1.6 8 40330 0
0.32 1.6 8 40 330 0
0.32 1.6 8 40 330 0
0.32 1.6 8 40 330 0
0.32 1.6 8 40 330 0
0.32 1.6 8 40 330 0
2 4 6 8 10 12 14
NIM induced gene expression (n=3) HepG2 cells
CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32
Concentration [µM]
Fo ld c h a n ge 2 - Δ Δ C T
0 1.6 8 40 200
1000 0
1.6 8 40 200
1000 0
1.6 8 40 200
1000 0
1.6 8 40 200
1000 0
1.6 8 40 200
1000 0
1.6 8 40 200
1000 0
1.6 8 40 200 1000 0
1 2 3 4 5 6 7 8
PhB induced biomarker expression (n=3) HepG2 cells
Fo ld c h a n ge 2 - Δ Δ C T
0 0.01
0.06
0.32 1.6 35 0 0.01
0.06
0.32 1.6 35 0 0.01
0.06
0.32 1.6 35 0 0.01
0.06
0.32 1.6 35 0 0.01
0.06
0.32 1.6 35 0 0.01
0.06
0.32 1.6 35 0 0.01
0.06
0.32 1.6 35 0
1 2 3 4 5 6
PMZ induced biomarker expression (n=3) HepG2 cells
CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32
Concentration [µM]
Fo ld c h a n ge 2 - Δ Δ C T
0 1 2 3 4 5 6 7
PPL induced biomarker expression (n=3) HepG2 cells
Fo ld c h a n ge 2 - Δ Δ C T
0
0.32 1.6 8 40200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40 200 0
0.32 1.6 8 40200 0
0.32 1.6 8 40 200 0
0.5 1 1.5 2 2.5 3 3.5 4
RIF induced biomarker expression (n=3) HepG2 cells
CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32 Baseline
Concentration [µM]
Fo ld c h a n ge 2 - Δ Δ C T
0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0 0.01
0.04 0.2 1 5 0
5 10 15 20 25