Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: Expression levels of FABP4 in sheep endometrium. (a) FABP4 mRNA expression in sheep endometrium on day 4 and day 15. (b, c) FABP4 protein expression in sheep endometrium on day 4 and 15. The optical density was normalized to the density of β-actin in the same lane. (d, e) Rabbit IgG group was used as the negative control for analyzing the localization and expression of FABP4 in sheep endometrium. Data are presented as mean ± standard error with significant differences at P < 0.05 and extremely significant differences at P < 0.01. * indicates P < 0.05, ** indicates P < 0.01, and no sign indicates that the difference is not significant. The technique was repeated thrice.

Article Snippet: SEECs were fixed with 4% paraformaldehyde at room temperature, blocked with BSA for 30 min, and FABP4 rabbit polyclonal antibody (Boster, BM4029, 1:50) and CK18 mouse monoclonal antibody (Abcam, Cambridge, UK, ab668, 1:50) were added overnight at 4°C.

Techniques: Expressing, Negative Control

Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: Expression of FABP4 in SEECs. (a) Isolation and purification of SEECs (250 ×). When the cells reached the sixth passage, they became larger and rounder and began to senesce. (b) Immunofluorescence identification of SEECs (CK18) (100 ×). (c) Localization of FABP4 in SEECs (Yellow arrows are FABP4 in the nuclei) (25 ×).

Article Snippet: SEECs were fixed with 4% paraformaldehyde at room temperature, blocked with BSA for 30 min, and FABP4 rabbit polyclonal antibody (Boster, BM4029, 1:50) and CK18 mouse monoclonal antibody (Abcam, Cambridge, UK, ab668, 1:50) were added overnight at 4°C.

Techniques: Expressing, Isolation, Purification, Immunofluorescence

Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: Hormone treatment of SEECs and detection of changes in FABP4 expression. (a) Protein expression levels of PGR and ER after combined hormone treatment. (b) The mRNA expression levels of ISG15, HOXA10, CXCL10, and RSAD2 after hormone treatment. (c) Levels of the prostaglandin secreted from the SEECs after hormone treatment.(d) Expression levels of FABP4 after hormone treatment. All data are presented as mean ± standard error. Differences were considered significant at P < 0.05 and extremely significant at P < 0.01.

Article Snippet: SEECs were fixed with 4% paraformaldehyde at room temperature, blocked with BSA for 30 min, and FABP4 rabbit polyclonal antibody (Boster, BM4029, 1:50) and CK18 mouse monoclonal antibody (Abcam, Cambridge, UK, ab668, 1:50) were added overnight at 4°C.

Techniques: Expressing

Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: FABP4 inhibition impedes SEEC function. (a, b) Mobility of SEECs at 0, 24, 48, 72 and 96 h was measured using a scratch test. Migratory capacity was calculated as a percentage of healing area relative to time 0. (c) CCK-8 viable cell counts quantified the proliferative capacity of SEECs treated with hormone and FABP4 inhibitor BMS309403. (d–g) Expression levels of EMT after hormone and inhibitor treatment (E-cadherin, N-cadherin, Vim, and β-catenin). (h, i) Changes in endoplasmic reticulum stress-related protein CHOP and GRP78 were measured. (j, k) Changes in autophagy-related proteins p-mTOR, LC3B II/I, and P62. Data are expressed as mean ± standard error. Differences were considered significant at P < 0.05 and extremely significant at P < 0.01.

Article Snippet: SEECs were fixed with 4% paraformaldehyde at room temperature, blocked with BSA for 30 min, and FABP4 rabbit polyclonal antibody (Boster, BM4029, 1:50) and CK18 mouse monoclonal antibody (Abcam, Cambridge, UK, ab668, 1:50) were added overnight at 4°C.

Techniques: Inhibition, CCK-8 Assay, Expressing

Journal: The Journal of Reproduction and Development

Article Title: FABP4 mediates endoplasmic reticulum stress and autophagy to regulate endometrial epithelial cell function during early sheep gestation

doi: 10.1262/jrd.2023-015

Figure Lengend Snippet: TG and 3-MA treatment partially restores SEEC function after BMS30940 suppression of FABP4. (a, b) Expression levels of key proteins in the endoplasmic reticulum stress signaling pathway after combined treatment with hormones, BMS309403, and TG. (c, d) Expression levels of key proteins in autophagy and apoptosis signaling pathways after combined treatment with hormones, BMS309403, and 3-MA. (e) Secreted prostaglandin levels from SEECs after combined treatment with hormones, BMS309403, and TG or 3-MA. Data are expressed as mean ± standard error. Differences were considered significant at P < 0.05 and extremely significant at P < 0.01.

Article Snippet: SEECs were fixed with 4% paraformaldehyde at room temperature, blocked with BSA for 30 min, and FABP4 rabbit polyclonal antibody (Boster, BM4029, 1:50) and CK18 mouse monoclonal antibody (Abcam, Cambridge, UK, ab668, 1:50) were added overnight at 4°C.

Techniques: Expressing, Protein-Protein interactions

Journal: Histochemistry and Cell Biology

Article Title: Expression and enhancement of FABP4 in septoclasts of the growth plate in FABP5-deficient mouse tibiae

doi: 10.1007/s00418-020-01953-y

Figure Lengend Snippet: FABP4-immnopositive cells in the chondro-osseous junction (COJ) of WT and FABP5 −/− mice. Light micrograph ( a ) and 3D-reconstructed image ( b , c ) of FABP5-immunopositive septoclasts in WT mice. Light micrograph ( d ) and 3D-reconstructed image ( e , f ) of FABP4-immunopositive cells in WT mice. Light micrograph ( g ) and 3D-reconstructed image ( h , i ) of FABP4-immunopositive cells in FABP5 −/− mice. d , g Arrowheads: FABP4 immunopositive cells. c , f , i Images drawing the outline of septoclasts as same as ( b , e and h ), respectively. Asterisks: cell bodies of septoclasts, Dotted lines: boundaries of septoclastic processes, Solid lines: outline of septoclastic body and processes, P length of process. j Mean SD, [ n = 16]. NS no significance. Scale bars: 100 μm ( a , d , g ), 6.1 μm (length of the side of a square in 3D grid, b , c , e , f , h , i )

Article Snippet: For double immunofluorescent staining, sections were treated overnight at room temperature with a mixture of rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech, Chicago, IL, USA) and goat anti-mouse CD31 (AF3628; 10 μg/ml; R&D system), or rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech) and goat anti-mouse PDGFRβ (AF1042; 10 μg/ml; R&D system), or rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech) and goat anti-human cathepsin B (AF965; 10 μg/ml, R&D Systems), or rabbit anti-rat FABP5 (0.5 μg/ml, Owada et al. ) and goat anti-mouse FABP4 (AF1443; 10 μg/ml, R&D Systems), or rabbit anti-human PPARγ (MA5-14,889; 1:50, Invitrogen, Carlsbad, CA, USA) and goat anti-mouse FABP4 (AF1443; 10 μg/ml, R&D Systems).

Techniques:

Journal: Histochemistry and Cell Biology

Article Title: Expression and enhancement of FABP4 in septoclasts of the growth plate in FABP5-deficient mouse tibiae

doi: 10.1007/s00418-020-01953-y

Figure Lengend Snippet: Expression of FABP4 in FABP5-immunopositive septoclasts in WT mice. Light micrographs of longitudinal sections at the chondro-osseous junction (COJ) of proximal tibiae of WT mice stained for FABP4 (red) plus cathepsin B (green) ( a ), for FABP4 (red) plus FABP5 (green) ( b ), for FABP4 (red) plus PDGFRβ (green) ( c ), and for FABP4 (red) plus CD31 (green) ( d ). Scale bars: 20 μm

Article Snippet: For double immunofluorescent staining, sections were treated overnight at room temperature with a mixture of rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech, Chicago, IL, USA) and goat anti-mouse CD31 (AF3628; 10 μg/ml; R&D system), or rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech) and goat anti-mouse PDGFRβ (AF1042; 10 μg/ml; R&D system), or rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech) and goat anti-human cathepsin B (AF965; 10 μg/ml, R&D Systems), or rabbit anti-rat FABP5 (0.5 μg/ml, Owada et al. ) and goat anti-mouse FABP4 (AF1443; 10 μg/ml, R&D Systems), or rabbit anti-human PPARγ (MA5-14,889; 1:50, Invitrogen, Carlsbad, CA, USA) and goat anti-mouse FABP4 (AF1443; 10 μg/ml, R&D Systems).

Techniques: Expressing, Staining

Journal: Histochemistry and Cell Biology

Article Title: Expression and enhancement of FABP4 in septoclasts of the growth plate in FABP5-deficient mouse tibiae

doi: 10.1007/s00418-020-01953-y

Figure Lengend Snippet: Cell count of FABP4-immunopositive septoclasts ( a ) and light micrographs of longitudinal sections stained for PPAR (red) plus FABP4 (green) with DAPI (blue) at the COJ of proximal tibiae of WT mice ( b – d ) and FABP5 −/− mice ( e – g ). Number of FABP5-immunopositive cells represents that of septoclasts ( a ). Arrowheads: lack ( b – d ) or occurrence ( e – g ) of immunoreactivity of PPAR in the nucleus of FABP4-positive septoclasts. a Mean SD, * P < 0.001 [ n = 20]. Scale bars: 20 μm

Article Snippet: For double immunofluorescent staining, sections were treated overnight at room temperature with a mixture of rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech, Chicago, IL, USA) and goat anti-mouse CD31 (AF3628; 10 μg/ml; R&D system), or rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech) and goat anti-mouse PDGFRβ (AF1042; 10 μg/ml; R&D system), or rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech) and goat anti-human cathepsin B (AF965; 10 μg/ml, R&D Systems), or rabbit anti-rat FABP5 (0.5 μg/ml, Owada et al. ) and goat anti-mouse FABP4 (AF1443; 10 μg/ml, R&D Systems), or rabbit anti-human PPARγ (MA5-14,889; 1:50, Invitrogen, Carlsbad, CA, USA) and goat anti-mouse FABP4 (AF1443; 10 μg/ml, R&D Systems).

Techniques: Cell Counting, Staining

Journal: Histochemistry and Cell Biology

Article Title: Expression and enhancement of FABP4 in septoclasts of the growth plate in FABP5-deficient mouse tibiae

doi: 10.1007/s00418-020-01953-y

Figure Lengend Snippet: Cell count of FABP4-immunopositive septoclasts ( a ) and light micrographs of longitudinal sections stained for PPAR (red) plus FABP4 (green) with DAPI (blue) at the COJ of proximal tibiae of control mice ( b – d ) and PPAR agonist (GW1929)-treated mice ( e – g ). Number of FABP5-immunopositive cells represents the number of septoclasts ( a ). Arrowheads: lack ( b – d ) or occurrence ( e – g ) of immunoreactivity of PPAR in the nucleus of FABP4-positive septoclasts. a Mean SD, * P < 0.01 [ n = 20]. Scale bars: 20 μm

Article Snippet: For double immunofluorescent staining, sections were treated overnight at room temperature with a mixture of rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech, Chicago, IL, USA) and goat anti-mouse CD31 (AF3628; 10 μg/ml; R&D system), or rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech) and goat anti-mouse PDGFRβ (AF1042; 10 μg/ml; R&D system), or rabbit anti-FABP4 (12,802–1-AP; 1:50, Proteintech) and goat anti-human cathepsin B (AF965; 10 μg/ml, R&D Systems), or rabbit anti-rat FABP5 (0.5 μg/ml, Owada et al. ) and goat anti-mouse FABP4 (AF1443; 10 μg/ml, R&D Systems), or rabbit anti-human PPARγ (MA5-14,889; 1:50, Invitrogen, Carlsbad, CA, USA) and goat anti-mouse FABP4 (AF1443; 10 μg/ml, R&D Systems).

Techniques: Cell Counting, Staining