Fig. 4

C-Kit⁺-LSECs alleviate NASH in vitro. A, B CD31⁺C-Kit⁺-pLSECs isolated from Con and MCD mice were detected by flow cytometry. C–G pHCs or pHSCs were cocultured with PA-treated C-Kit⁺- or C-Kit⁻-pLSECs. ORO staining C and calculation D of lipid droplets in pHCs (× 400). IF staining C and calculation E of TNF-α (green IF) in PHCs (× 400). IF staining C and calculation F of α-SMA (red IF) in pHSCs (× 400). DAPI (blue) was used for nuclear staining. (G) The mRNA of TNF-α (in pHCs) and α-SMA (in pHSCs) was detected by qPCR. (H-M) HepG2 or LX2 cells were cocultured with 6 groups of TMNK-1 cells (BSA, PA, sh-NC + PA, sh-C-Kit + PA, ov-NC + PA, ov-C-Kit + PA). ORO staining H and calculation I in HepG2 cells (× 1000). IF staining () and calculation J of TNF-α (red IF) in HepG2 cells (× 500). IF staining H and calculation K of α-SMA (red IF) in LX2 cells (× 200). DAPI (blue) was used for nuclear staining. The mRNA of L lipid metabolism genes (APDN, FXR, PPAR-α, and LXR) and M inflammation and fibrosis genes (TNF-α, IL-6, Col1a, and α-SMA) was examined by qPCR. The p-value indicated statistical significance compared to the pLSEC-Con, C-Kit⁻-pLSEC, BSA, sh-NC + PA or ov-NC + PA group