1. Effects of declining sorafenib levels and dose on acquired reversible resistance and toxicity in hepatocellular carcinoma Elizabeth A. Kuczynski, Christina R. Lee, Shan Man, Eric Chen, Robert S. Kerbel SUPPLEMENTARY FIGURES S1-5

2. C Hep3B-hCG parental Hep3B-hCG- R1 Hep3B-hCG- R2 D A B EGF Sorafenib P-ERK ERK ParentalR1 -+ - + - + - + - - + + E F G Invasion Proliferation Tumor growth (hCG) Clonogenic Survival hCG: Orthotopic Transfer hCG: Subcutaneous Transfer Supplementary Figure S1 IC50: 3.03µM (P), 2.92µM (R1), 2.36µM (R2)

3. Supplementary Figure S2 B hCG for Drug Level Assessment A PlasmaTumor *

4. hCG: 65 Day Sorafenib Pre- Conditioned Mice A B Supplementary Figure S3 +2d implant, +5d treat hCG: Indicator Group of SCID HCC mice p=0.742 p=0.965 P=0.0659 C

5. Supplementary Figure S4 Weight Loss: HCC vs. Tumor-Free F G p=0.266 Survival: HCC vs. Tumor-Free D E ** Tumor + Liver Weight p=0.600 Survival: Dose escalation (30 mg/kg) p=0.165 p=0.285 B hCG: Dose Escalation Early vs. Late Progressors *p=0.0145 Sorafenib concentrations: day 14 C p=0.202 p=0.152 p=0.00554 A

6. A C Liver Lysates - Tumor-Free Control* Sensitive Resistant Escalated Liver lysates - HCC SensitiveResistantEscalatedControl P-STAT3 STAT3 P-STAT3 STAT3 Supplementary Figure S5 D ControlSensitiveResistantEscalated CAIX Actin ControlSensitive Resistant Escalated P-PDGFRβ PDGFRβ Tumor Lysates B P-ERK ERK