Journal: bioRxiv
Article Title: Estrogen Receptor 1 Signaling in Hepatic Stellate Cells Designates Resistance to Liver Fibrosis
doi: 10.1101/2024.10.10.617541
Figure Lengend Snippet: (A) HSCs in the liver tissues of Lrat Cre ; RCL-ChR2(H134R)/EYFP +/- mice fed with the normal chow diet were examined by FACS. A vast majority (>99%) of retinol + CD45 - HSCs were EYFP-positive. (B to J) Lrat Cre ;Esr1 fl/fl or control Esr1 fl/fl female littermates of 8 weeks old received the normal chow diet (NCD) or methionine-choline deficient (MCD) diet to induce liver fibrosis. (B) HSCs were FACS-sorted from the liver tissues of Lrat Cre ;Esr1 fl/fl or control Esr1 fl/fl mice. Genetic deletion of Esr1 in Lrat Cre ;Esr1 fl/fl HSCs was validated by the qPCR analysis. mean ± SEM, ** p < 0.01 (Student’s t -test). (C to G) Paraffin sections of the liver tissues of indicated conditions were assessed by histochemistry or immunohistochemistry. (C) Representative images of H&E staining. (D and E) Representative images of Sirius Red staining (D) and the quantification of the percentage (%) of Sirius Red-positive area (E) . (F and G) Representative images of anti-α-SMA immunohistochemistry (F , black arrows exemplify anti-α-SMA signals ) and the quantification of the percentage (%) of α-SMA-positive area (G) . mean ± SEM, * p < 0.05, ** p < 0.01 (Student’s t -test). (H and I) Cryosections of the liver tissues were examined by the immunofluorescence co-staining of TIMP1 and α-SMA. (H) Representative images were shown. White arrowheads exemplify anti-TIMP1 signals. (HI The mean fluorescence intensity of anti-TIMP1 signals was quantified. mean ± SEM, * p < 0.05 (Student’s t -test). (I) mRNA levels of fibrosis-related genes were determined by the qPCR analyses. mean ± SEM, * p < 0.05, n.s., not significant (two-way ANOVA test).
Article Snippet: For the immunofluorescence staining, 10-μm cryosections were blocked with PBS / 3% BSA and immunostained with the intended primary antibodies, including rabbit anti-ESR1 antibody (Millipore, #06-935), rabbit anti-ESR2 antibody (Invitrogen, #PA1311), goat anti-α-SMA antibody (Novus, #NB300-978), rabbit anti-TIMP1 antibody (Bioss, #bs-0415R), and chicken anti-GFP antibody (Aves Labs, #GFP-1010).
Techniques: Control, Immunohistochemistry, Staining, Immunofluorescence, Fluorescence