Figure 5.
AAV-Mediated Overexpression of HOXB6 Suppresses the Proliferation and Differentiation of SOX9+ LPCs to Aggravate Damage in a CCl4 Chronic Liver Injury Model
AAV-ctrl and AAV-Hoxb6 were administered to 8-week-old C57 mice via tail vein injection. After 2 weeks, mice were treated with CCl4 (diluted in oil with 1:4) twice per week for 4 weeks, then livers and blood were collected.
(A) Hepatic expression levels of SOX9 and HOXB6 were measured in AAV-ctrl, AAV-HOXB6 mice after CCl4 injury by qRT-PCR and western blot.
(B) Representative H&E, Masson, and Sirius red staining in AAV-ctrl mice liver and AAV-Hoxb6 mice liver after chronic hepatic injury by CCl4 treatment. Fibrosis was quantified by morphometric measurement of Masson and Sirius red.
(C) Serum ALT, AST, and LDH levels of AAV-ctrl mice and AAV-HOXB6 mice after chronic hepatic injury by CCl4 treatment.
(D) Representative images from RNAscope assays of Hnf4α and Sox9 in livers of AAV-ctrl and AAV-Hoxb6 mice after CCl4 injury. Red presents Sox9. Blue (DAPI) shows nuclei. Green (Hnf4α) marks hepatocytes. Scale bar represents 20 μm.
(E) SOX9 and HNF4α double staining in periportal areas in the livers of AAV-ctrl and AAV-Hoxb6 mice after CCl4 injury. Blue (DAPI) shows nuclei. Red (HNF4α) marks hepatocytes. Green presents SOX9. Scale bar represents 20 μm.
(F) SOX9 and BrdU double staining in periportal areas in livers of AAV-ctrl and AAV-Hoxb6 mice after CCl4 injury. Blue (DAPI) shows nuclei. Red (BrdU) marks the proliferating cells. Green presents SOX9. Scale bar represents 20 μm.
Data are expressed as means ± SD, n = 6 mice per group containing three replicates. Significant difference is presented at the levels of ∗p < 0.05 and ∗∗p < 0.01 by two-tailed Student's t test. See also Figure S4.