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: C57BL/6 wild-type female mice of 6 weeks old underwent ovariectomy or sham surgery. The mice were then fed with the normal chow diet (NCD) or methionine-choline deficient (MCD) diet to induce liver fibrosis. (A) Diagram of the experimental procedure. (B to F) Paraffin sections of the liver tissues of indicated conditions were assessed by histochemistry or immunohistochemistry. (B) Representative images of H&E staining. (C and D) Representative images of Sirius Red staining (C) and the quantification of the percentage (%) of Sirius Red-positive area (D) . (E and F) Representative images of anti-α-SMA immunohistochemistry (E , black arrows exemplify anti-α-SMA signals ) and the quantification of the percentage (%) of α-SMA-positive area (F) . mean ± SEM, * p < 0.05, ** p < 0.01 (Student’s t -test). (G and H) Cryosections of the liver tissues were examined by the immunofluorescence co-staining of TIMP1 and α-SMA. (G) Representative images were shown. White arrowheads exemplify anti-TIMP1 signals. (H) 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: Immunohistochemistry, Staining, Immunofluorescence, Fluorescence

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: C57BL/6 wild-type male mice of 4 weeks old underwent castration and 17β-estradiol (β-E2) hormone replacement. The mice then received the normal chow diet (NCD) or methionine-choline deficient (MCD) diet to induce liver fibrosis. (A) Diagram of the experimental procedure. (B to F) Paraffin sections of the liver tissues of indicated conditions were assessed by histochemistry or immunohistochemistry. (B) Representative images of H&E staining. (C and D) Representative images of Sirius Red staining (C) and the quantification of the percentage (%) of Sirius Red-positive area (D) . (E and F) Representative images of anti-α-SMA immunohistochemistry (E , black arrows exemplify anti-α-SMA signals ) and the quantification of the percentage (%) of α-SMA-positive area (F) . mean ± SEM, * p < 0.05, ** p < 0.01 (Student’s t -test). (G and H) Cryosections of the liver tissues were examined by the immunofluorescence co-staining of TIMP1 and α-SMA. (G) Representative images were shown. White arrowheads exemplify anti-TIMP1 signals. (H) 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 < 0.01, 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: Immunohistochemistry, Staining, Immunofluorescence, Fluorescence

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