Supplemental Figure S1. Purity of Parenchymal and Non-Parenchymal Cell Isolations. (A

Supplemental Figure S1. Purity of parenchymal and non-parenchymal cell isolations. (A and C) Mice were injected with 3 doses of NanoCurc™ (equivalent to 25 mg/kg body weight of free curcumin) in a 24 hour period. Parenchymal (hepatocytes) and non-parenchymal cells (NPC) were isolated by digesting the liver with collagenase in situ. The isolated NPCs (A) and hepatocytes (C) were smeared onto slides, fixed and stained with H&E histochemical stain. Hepatocytes in NPC fraction are indicated with a yellow arrow (A) and NPCs in the hepatocyte fraction are indicated with a black arrow(C). Cells containing cytoplasmic fat droplets in the NPC fraction are shown with green arrow (A).200 cells (NPC and hepatocytes) were counted from the H&E stained slide at 40 X resolution to determine the purity of the NPC (B)and hepatocytes (D) fractions. Percentage purity was calculated by dividing the number of NPC or hepatocytes by 200 and multiplying by 100.

Supplemental Figure S2. Effect of NanoCurc™ on Innate and Adaptive Immune Cells.The mice were untreated (PBS) or injected with either void nanoparticles (Poly) or NanoCurc™ (NC) along with CCl4 according to the treatment protocol in Materials and Methods. Livers harvested 24 hours after the last NanoCurc™ treatment; and liver leukocytes were isolated and stained with the indicated cell markers. Representative flow cytometry dot plots and histograms from of two experiments with mean and standard error of the mean.

Supplemental Figure S3. Cells were seeded in a 24-well plate and allowed to attach over night. Then the cell were treated for 20 hours with varying concentrations (5, 10, 15, 20 μM) of free curcumin dissolved in 0.1 % DMSO or the equivalent amount of NanoCurc™ dissolved in sterile pyrogen free water. After paraformaldehyde fixation the cells were incubated with cleaved caspase-3 and then Alexa Fluor 568 labeled with bright orange red fluorescent dye. After washing and counter staining with DAPI, the cells were observed under the fluorescent microscope. (A) DAPI stains all the nuclei blue whereas the anti cleaved caspase 3 (CC3) stains apoptotic cells red. (B) DAPI positive and CC3 positive cells were counted at 40X in 10 fields. Percentage CC3 to DAPI was calculated by dividing number of CC3 positive cells by total number of cells multiplied by 100.