APAP induced biomarker expression (n=3) PHH

(1)

Figure S3: Compound induced biomarker expression in primary human hepatocytes. The presented values (relative expression fold changes) represent mean values that were calculated from 1-3 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

0.5 1 1.5 2 2.5 3 3.5 4 4.5 5

APAP induced biomarker expression (n=3) PHH

CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32

Concentration [mM]

Fo ld c h an ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for 1 donor at 8 µM and 40 µM.

(2)

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 3.5

ASP induced biomarker expression (n=2) PHH

Concentration [mM]

Fo ld c h a n ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for 1 donor at all concentrations.

(3)

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 3.5 4

BPR induced biomarker expression (n=2) PHH

Concentration [nM]

Fo ld c h a n ge 2 - Δ Δ C T

(4)

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

CBZ induced biomarker expression (n=2) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

(5)

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

1 2 3 4 5 6

CHL induced biomarker expression (n=2) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for 1 donor at 2.6 nM and 64 nM.

(6)

(7)

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

5 10 15 20 25 30 35 40 45

DFN induced biomarker expression (n=2) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

(8)

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=2) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for 1 donor at all concentrations.

(9)

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

1 2 3 4 5 6

HYZ induced biomarker expression (n=1) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

(10)

0 10 20 30 40 50 60 70

INAH induced biomarker expression (n=2) PHH

Concentration [mM]

Fo ld c h a n ge 2 - Δ Δ C T

(11)

(12)

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=1) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for this donor.

(13)

0

0.32 1.6 8 40200 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=2) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for 1 donor at 200 µM.

(14)

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=1) PHH

Concentration [nM]

Fo ld c h a n ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for this donor at 20 pM, 0.51 nM, 2.6 nM and

100 nM.

(15)

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

2 4 6 8 10 12 14

NFT induced biomarker expression (n=2) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for 1 donor at all concentrations.

(16)

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

2 4 6 8 10 12

NIM induced gene expression (n=1) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for this donor at all concentrations.

(17)

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

PhB induced biomarker expression (n=2) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

(18)

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=2) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

SULT1C2 expression was below the detection level for 1 donor at 320 nM.

(19)

0 0.5 1 1.5 2 2.5 3

PPL induced biomarker expression (n=1) PHH

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

(20)

0

0.32 1.6 8 40 200 0

1 2 3 4 5 6 7 8

RIF induced biomarker expression (n=3) PHH

CYP1B1 CYP3A7 Tubb2b Fibin G6PD RGCC Fbxo32 Baseline

Concentration [µM]

Fo ld c h a n ge 2 - Δ Δ C T

(21)

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 30 35 40

VPA induced biomarker expression (n=3) PHH

CYP1B1 CYP3A7 Tubb2b SULT1C2 Fibin G6PD RGCC Fbxo32 Threshhold