FIGURE LEGENDS
Figure 1. Tenofovir alafenamide (TAF) treatment ameliorates
liver injury in the nonalcoholic steatohepatitis (NASH) mouse model.(A) Experimental schedule for the NASH mouse model using streptozotocin
(STZ) injection and high-fat, high-cholesterol (HFHC) diet. C57BL/6J
mice were injected with 0.2 mg STZ and fed HFHC diet or carbohydrate
(CHO) diet and treated with mock or TAF. NASH resulted from the HFHC
diet after STZ subcutaneous injection for 4 weeks (n = 6-10). (B) Serum
alanine aminotransferase (ALT), aspartate aminotransferase (AST), and
triglyceride (TRIG) levels in STZ-injected, HFHC-fed NASH experimental
animals treated with mock or TAF. (C) Representative histological
analyses of liver sections stained with haematoxylin and eosin, Sirius
red, anti-lipopolysaccharide (LPS) and anti-toll-like receptor 4 (TLR4)
expression in STZ-injected mock-treated HFHC diet-fed and STZ-injected
TAF-treated HFHC diet-fed mice livers. Original magnification: 50×. (D)
Comparison of the frequency of LPS- or TLR4-positive cells between
STZ-injected, HFHC diet-fed NASH experimental animals treated with mock
or TAF. Morphometric analysis of Sirius Red-stained liver section was
performed from 5 fields of 5 liver sections per group. Comparison of
NASH Activity Score (NAS) from mouse liver specimens among each
treatment group. (E) Experimental schedule for the NASH mouse model
using choline-deficient, L-amino acid-defined, high-fat (CDAHF) diet-fed
mock or TAF. C57BL/6J mice were fed a CDAHF or CHO diet for 6 weeks (n =
7-10). (F) Serum ALT, AST, and TRIG levels in CDAHF diet-fed NASH
experimental animals treated with mock or TAF. *p<0.05, **p <0.001, ***p<0.001.
Figure 2. TAF decreases the number of activated mononuclear
phagocytes (MPs) in the STZ-injected, HFHC diet-fed NASH mouse liver.(A) Representative dot plot describing the proportions of
recruited/resident MPs. Data are representative of five independent
experiments. (B) Recruited (CD11bhighF4/80low) and resident (CD11blowF4/80high) MP cell number per g liver weight and cell
percentage in the mock or TAF-treated STZ-injected, HFHC diet-fed NASH
mouse model was counted using flow cytometry. (C) Mean fluorescence
intensity (MFI) of programmed death ligand 1 (PD-L1), IAIE and MER
proto-oncogene, and tyrosine kinase (MerTK) in CD11b+F4/80+ cells. (D) Frequency of
PD-L1+/IAIE+/MerTK+intrahepatic MP cells in the STZ-injected, HFHC diet-fed NASH mouse
model treated with mock or TAF. (E) Relative mRNA expression of
inflammatory genes in mock or TAF-treated NASH mouse liver. *p<0.05, **p <0.001, ***p<0.001.
Figure 3. TAF decreases the number of activated MPs in the
choline-deficient, L-amino acid-defined, high-fat (CDAHF) diet-fed NASH
mouse liver. (A) Representative dot plot describing the proportions of
recruited/resident MPs. Data are representative of five independent
experiments. (B) Recruited (CD11bhighF4/80low) and resident (CD11blowF4/80high) MP cell number per g liver weight and cell
percentage in the mock or TAF-treated CDAHF diet-fed NASH mouse model
was counted using flow cytometry. (C) MFI of PD-L1, IAIE and MerTK in
CD11b+ F4/80+ cells. (D) Frequency
of
PD-L1+/IAIE+/MerTK+intrahepatic MP cells in the CDAHF diet-fed NASH mouse model treated
with mock or TAF. *p <0.05, **p<0.001, ***p <0.001.
Figure 4. Ex vivo effect of TAF on MPs isolated from the
livers of the thioacetamide (TAA)-induced liver injury mouse model. (A)
The liver injury mouse model experimental schedule using TAA injection
(n = 5). Mice were intraperitoneally injected with 100 or 150 mg/kg TAA
three times a week. (B) Representative histological analyses of liver
sections stained with H&E, Sirius red in TAA-treated mouse liver.
Original magnification: 50×. Morphometric analysis of Sirius Red-stained
liver section was done from 5 fields of 5 liver sections per group. (C)Ex vivo experimental scheme of MPs isolated from the liver injury
mouse model treated with mock or TAF. The mice livers were perfused, and
MPs were isolated using gradient centrifugation. (D) Frequency of
PD-L1+ cells among isolated
CD11b+F4/80+ cells after treatment
with mock or TAF. (E) MFI of PD-L1 in isolated
CD11b+F4/80+ cells after mock or TAF
treatment. *p <0.05, **p <0.01,
***p <0.001.