Fig 2: FA improves CCl4-induced oxidative stress and hepatic injury in MPHs. (A) A CCK-8 assay. (B, C, F and G) MPHs were pre-treated with FA at different concentrations for 1 h and then treated with CCl4 (10 mM) for another 24 h (B) MDA and SOD levels in MPHs. (C) Representative images of immunofluorescent staining of ALB in MPHs. (D) MPHs were treated with 25 µM FA at different time points or different concentrations of FA for 24 h. (E) The pathway of AMPK and NOX2 involved in oxidative stress. (G) Representative images of ROS immunofluorescent staining in MPHs. (H) After pre-treated with CC (2 µM) or FA (25 µM) or both, MPHs were administrated with CCl4 (10 mM) for another 2 h or 24 h (D, F and H) Representative immunoblots against p-AMPK, t-AMPK, p-LKB1, t-LKB1, p-ERK1/2, t-ERK1/2, NOX2 and ß-ACTIN were shown. Statistical significance: **p < 0.01, ***p < 0.001, compared with control group; # p < 0.05, ## p < 0.01, ### p < 0.001, compared with CCl4 group. One-way ANOVA with Tukey’s post-hoc tests (n = 3).

Fig 3: The level of FcRn expression regulates the intracellular levels of albumin in tumor cellsA. HCC1419 cells transduced with empty vector (EV), scrambled shRNA (scramble) or shRNAs targeting FcRn (sh-5, sh-6) were pulsed with 1 µg/ml Alexa 647-labeled FcRn-specific nanobody for 40 minutes at 37°C in medium (pH 6.0). Alexa 647 levels (MFI) were determined using flow cytometry and mean values for triplicate samples are shown. Control indicates autofluorescence levels of cells. The right hand panel shows immunoblotting of cell lysates using antibodies specific for FcRn a-chain and ß-actin. B. DU145 cell lines transduced with empty vector (EV) and FcRn expression constructs (WT-FcRn or H166A-FcRn) were analyzed as in panel (A). C, D. HCC1419 cell lines (same as in panel A) were pulsed with 1.5 µM (100 µg/ml) MSA (C) or 1.5 µM (100 µg/ml) HSA (D) for 0 h or 2 hrs at 37°C at pH 7.4. Cell lysates were used in immunoblotting with antibodies specific for albumin (MSA/HSA), FcRn a-chain and ß-actin. Albumin levels were quantitated and normalized relative to HCC1419 cells. E, F. DU145 cell lines (same as in panel B) were treated with MSA (E) or HSA (F) as in panels (C) and (D), and albumin levels quantitated and normalized relative to DU145 cells. G. DU145 cell lines were pulsed with 1.5 µM Alexa 647-labeled MSA for 2 hrs. Cells were chased for the indicated times. Alexa 647 levels (MFI) were determined using flow cytometry and mean values for triplicate samples are shown (left panel). The right hand panel shows the MFI values normalized to the no chase (0 time point) MFI value for each cell line. H. HCC1419 cell lines transduced with scrambled shRNA (scramble), sh-5 or sh-5 plus shRNA resistant FcRn (Res-FcRn) were treated and albumin levels relative to ‘scramble’ analyzed as in panel (C). For panels (C-F) and (H), mean normalized signal levels derived from three independent immunoblotting experiments are shown. For the immunoblots, cropped images are shown with molecular weights (kDa) on the right. For panels (A-H), error bars represent S.D. Significant differences are indicated by * (one-way ANOVA, p < 0.05). For panels (A, B) and (G), data are representative of at least two independent experiments.