The number of
PD-L1+/IAIE+/MerTK+intrahepatic MPs is lower in TAF-treated STZ, HFHC diet-fed NASH mice.
We performed flow cytometry analysis after mouse liver digestion to
confirm the role of intrahepatic MPs in the NASH mouse model using the
livers of STZ and HFHC diet-fed mouse treated with mock or TAF. A gating
strategy for quantifying intrahepatic MPs using flow cytometry is shown
in Supplementary Figure S2A.
A representative gating strategy for intrahepatic recruited
(CD11bhigh F4/80low) and resident
(CD11blow F4/80high) MPs is shown in
Fig 2A. The TAF-treated HFHC diet-fed group showed significantly
decreased intrahepatic recruited (CD11bhighF4/80low) MPs than the mock-treated one. However,
there were no significant differences in resident
(CD11blow F4/80high) MPs (Fig 2B)
between the TAF-treated HFHC diet-fed group and the mock-treated one.
Programmed death ligand 1 (PD-L1), IAIE, MER proto-oncogene, and
tyrosine kinase (MerTK) were used as CD11b+F4/80+ MP activation surface markers. The intrahepatic
IAIE+ and MerTK+ MP mean
fluorescence intensity (MFI) values were significantly lower in the
TAF-treated HFHC diet group than in the mock-treated one (Fig 2C). The
intrahepatic
PD-L1+/IAIE+/MerTK+-positive
MP number was significantly decreased in the TAF-treated HFHC diet group
than in the mock-treated one (Fig 2D). Furthermore, the number of
CD11b+ F4/80+ MPs per g liver weight
was significantly reduced in the TAF-treated HFHC diet group than in the
mock-treated one (Supplementary Fig S2B). Figure 2E shows that tumour
necrosis factor-α, chemokine (C-C motif) ligand 2, and interleukin-1
beta mRNA expression levels were significantly lower in the TAF-treated
HFHC diet group than in the mock-treated one, indicating reduced
inflammation.