Anti β actin mab1501

HuH-7 cells were either left untreated or treated with rapamycin (Sigma), unformulated curcumin, or phytosomal curcumin with the indicated concentrations for 24 hours. Western blot analysis was performed as described^{62 (link)}. Briefly, total proteins were extracted with lysis buffer containing protease and phosphatase inhibitor cocktail (Roche Diagnostics, Mannheim, Germany) from cells or mice liver tissues after treatment. Equal amounts of proteins for each sample were resolved on sodium dodecyl sulfate- polyacrylamide gels and transferred to polyvinylidene difluoride membranes. Membranes were incubated with primary antibodies, followed by secondary antibodies, and then developed by an enhanced chemiluminescence system (Amersham Pharmacia Biotech, Amersham, UK). The primary antibodies used in this study were anti-mTOR (2972), anti-NF-κB p65 (8242), and anti-p-NF-κB p65 (Ser536) (3033) from Cell Signaling Technology (Danvers, MA, USA), anti-p-mTOR (Ser2448) (ab1093) from Abcam (Cambridge, UK), and anti-β-Actin (MAB1501) from Millipore (Billerica, MA, USA). β-Actin was used as the internal control. Full blots were shown in Fig. S5.

The following antibodies were used in this study: Anti-PrP antibodies 3F4 and 8H4 (Signet Laboratories, Dedham, MA), and SAF32 (189720, Cayman chemicals, USA), anti-ferritin (F5012, Sigma Aldrich, USA), anti-TfR (136800, Invitrogen, USA), anti-Tf (GTX2123, GeneTEX, USA), anti-ceruloplasmin (Dako, USA), Mouse IgG (5415, Cell signaling Technology, USA), anti-Fpn (NBP1-21502, Novus biological, USA), anti β-actin (MAB1501, Milipore, USA) and anti GAPDH (GT239, GeneTEX). HRP-conjugated anti-mouse, NA931V and anti-rabbit, NA934V (GE Healthcare, UK). Alexa Fluor-conjugated secondary antibodies (molecular probes), PNGase F (P0704S, NEB, USA). NAP (Non-animal protein) blocking solution (786-190T, G-Biosciences, USA), Hoechst (#33342, Invitrogen, USA), Fluoromount-G (Southern Biotech, USA).