bs 1630r bioss rabbit anti fabp4 polyclonal antibody  (Bioss)

Journal: eLife

Article Title: FABP4-mediated lipid accumulation and lipolysis in tumor-associated macrophages promote breast cancer metastasis

doi: 10.7554/eLife.101221

Figure Lengend Snippet:

Article Snippet: Antibody , Goat anti-mouseFABP4 polyclonal , R&D Systems , Cat#AF1443 , 1:300.

Techniques: Concentration Assay, Staining, Saline, SYBR Green Assay, Red Blood Cell Lysis, Cell Culture, XF Assay, Reverse Transcription, Polymer, Plasmid Preparation, Blocking Assay, Enzyme-linked Immunosorbent Assay, Sequencing, Transfection, Recombinant, Microarray, Expressing, Derivative Assay, Real-time Polymerase Chain Reaction, Software, Sterility, Migration, Pore Size, Flow Cytometry, Quantitative RT-PCR, Imaging, Microscopy, Transmission Assay, Electron Microscopy

Journal: EMBO Molecular Medicine

Article Title: FABP4 as a therapeutic host target controlling SARS-CoV-2 infection

doi: 10.1038/s44321-024-00188-x

Figure Lengend Snippet: ( A ) Immunohistochemical staining of lung biopsies of controls and COVID-19 patients, using anti-FABP4 antibody. ( B , C ) Circulating FABP4 concentrations measured from ( B ) female ( n = 109 female patients) and ( C ) male COVID-19 patients ( n = 174 male patients) and healthy controls ( n = 32 female and n = 13 male), stratified based on disease severity (**** p < 0.0001, ** p = 0.002, * p = 0.0221). ( D ) Circulating IL-6 levels in COVID-19 patients, stratified based on disease severity (** p = 0.0017). ( B – D ) Statistical analysis was performed using one-way ANOVA. ( E , F ) Effect size estimates and inference based on regression analysis of FABP4 concentration on ( E ) COVID-19 severity and ( F ) oxygen support measures while accounting for time of collection post symptom onset, age, sex, and BMI, using the linear mixed model to account for the patient-level random effects. ( E ) The healthy controls and, ( F ) patients who did not require oxygen support were used as a reference group. p-values are calculated based on the Wald test. For ( B – F ), the analysis included n = 283 total patients and n = 45 healthy controls. ( G , H ) FABP4 concentration in patients with severe and critical disease ( n = 176 patients), stratified based on ( G ) presence or absence of comorbidities (listed in Table and Dataset , *** p = 0.0009) or ( H ) age (*** p = 0.0006). Statistical analysis was performed using Welche’s t-test. ( B – H ) Data are derived from patient cohort 1 (collected November 2020–May 2021). Patients were sampled longitudinally, and the data shown in ( B ), ( C ), ( D ), ( G ) and ( H ) represent the maximum measured concentration per patient. Data are shown as the mean ± s.e.m. .

Article Snippet: The following antibodies were used for this study: anti-SARS-CoV-2 nucleocapsid mouse antibody (Cell Signaling Technology, 33717—1:200 dilution), anti-double stranded RNA J2 mouse antibody (Exalpha, 10010500—1:200 dilution), anti-calnexin rabbit antibody (Cell Signaling Technology, 2679—1:200 dilution), anti-FABP4 rabbit monoclonal antibody (Abcam, ab216708—1:200 dilution), anti-FABP4 goat polyclonal antibody (Novus, AF1443—1:200 dilution), anti-mouse secondary antibody (Cell Signaling Technology, 4410—1:1000 dilution), anti-rabbit secondary antibody (ThermoFisher Scientific, A-11037—1:1000 dilution).

Techniques: Immunohistochemical staining, Staining, Concentration Assay, Derivative Assay

Journal: EMBO Molecular Medicine

Article Title: FABP4 as a therapeutic host target controlling SARS-CoV-2 infection

doi: 10.1038/s44321-024-00188-x

Figure Lengend Snippet: ( A , B ) Maximum concentrations of circulating ( A ) FABP4 (**** p < 0.0001, * p = 0.044) and ( B ) IL-6 (*** p = 0.0005) of cohort 2 of COVID-19 patients ( n = 166) stratified based on disease severity (moderate: n = 52, severe: n = 21, and critical: n = 42). ( C ) Circulating levels of C-reactive protein (**** p < 0.0001, *** p = 0.0002, * p = 0.0221), ( D ) leukocytes (**** p < 0.0001, ** p = 0.0015) and ( E ) lymphocytes (**** p < 0.0001) of COVID-19 patients measured on the day in which the maximum FABP4 concentration was measured (day post symptom onset). Statistical analysis was performed using one-way ANOVA ( n = 283 cohort 1, and n = 116 cohort 2). ( F – I ) Maximum FABP4 concentration pooled from severe and critically ill patients (cohort 1: n = 176, cohort 2: n = 63), stratified based on ( F ) the presence or absence of comorbidities (listed in Table and Dataset , **** p < 0.0001), ( G ) age (** p = 0.0011), ( H ) BMI (*** p = 0.0001, ** p = 0.0098 and 0.004, * p 0.0263), and ( I ) the presence or absence of cardiometabolic conditions (diabetes, hypertension, or coronary artery disease, **** p < 0.0001). Statistical analysis for ( F ), ( G ) and ( I ) were performed using Welch’s t-test and one-way ANOVA for ( H ). Data are shown as the mean ± s.e.m. .

Article Snippet: The following antibodies were used for this study: anti-SARS-CoV-2 nucleocapsid mouse antibody (Cell Signaling Technology, 33717—1:200 dilution), anti-double stranded RNA J2 mouse antibody (Exalpha, 10010500—1:200 dilution), anti-calnexin rabbit antibody (Cell Signaling Technology, 2679—1:200 dilution), anti-FABP4 rabbit monoclonal antibody (Abcam, ab216708—1:200 dilution), anti-FABP4 goat polyclonal antibody (Novus, AF1443—1:200 dilution), anti-mouse secondary antibody (Cell Signaling Technology, 4410—1:1000 dilution), anti-rabbit secondary antibody (ThermoFisher Scientific, A-11037—1:1000 dilution).

Techniques: Concentration Assay

Journal: EMBO Molecular Medicine

Article Title: FABP4 as a therapeutic host target controlling SARS-CoV-2 infection

doi: 10.1038/s44321-024-00188-x

Figure Lengend Snippet: ( A – C ) hTERT pre-adipocytes and differentiated adipocytes infected with SARS-CoV-2 (WA1/2020, MOI = 1). ( A , B ) Relative expression of viral ( A ) genomic RNA (nucleocapsid, *** p = 0.0002, ** p = 0.0018) and ( B ) sub-genomic RNA (ORF1ab, *** p = 0.0003, * p = 0.0263), normalized to β-actin. ( C ) Viral loads measured from supernatant using plaque assay. (**** p < 0.0001, ** p = 0.0024). Data are pooled from two independent experiments ( n = 8, biological replicates). Statistical analysis was performed using two-way ANOVA. ( D ) IL-6 levels in the supernatant of differentiated adipocytes with or without viral infection (MOI = 1), measured by ELISA. Data are pooled from three independent experiments ( n = 14, biological replicates, **** p < 0.0001). Statistical analysis was performed using one-way ANOVA. ( E – G ) Adipocytes were infected at 4, 9, 12, 16, and 20 days post-differentiation (MOI = 1), with measurements taken 48 h post-infection. ( E ) Viral loads measured by plaque assay; data represent two independent experiments ( n = 4, biological replicates, **** p < 0.0001). Statistical analysis was performed using one-way ANOVA. ( F ) Western blot of viral nucleocapsid, FABP4, and β-actin proteins levels in cell lysates. ( G ) Quantification of nucleocapsid and FABP4 band intensities normalized to β-actin, representative of two independent experiments ( n = 4, biological replicates, **** p < 0.0001, *** p = 0.0001 and 0.0006). Statistical analysis was performed using two-way ANOVA. ( H , I ) Percent area of lipid droplets in infected cells and neighboring cells, quantified by fluorescence neutral lipid staining (Bodipy). Infected cells identified by ( H ) nucleocapsid-positive staining and ( I ) dsRNA-positive staining. Data pooled from two independent experiments ( n = 6, biological replicates, **** p < 0.0001, *** p = 0.0001); statistical analysis was performed using a standard t-test. ( J ) Representative confocal images of infected differentiated adipocytes (MOI = 1), stained for dsRNA (red), lipid droplets (yellow), and nucleus (DAPI, blue). Scale bar = 50 μm; magnified regions = 10 μm ( n = 3). ( K ) Percent lipid droplet area relative to dsRNA-positive area per cell. Pearson correlation coefficient indicated as r. ( L , M ) Representative confocal images of control and infected adipocytes at 8, 12, 24, and 48 h post-infection, stained for dsRNA (red), lipid droplets (yellow), calnexin (blue), and FABP4 (gray). ( L ) Merged image of all stains (Scale bar = 50 μm); insets highlight overlap of FABP4, dsRNA, and calnexin (Scale bar = 5 μm). ( M ) Signal overlap between dsRNA, FABP4, and calnexin, and between FABP4 and calnexin individually (Scale bar = 5 μm). ( N – P ) Colocalization of target signals over time, represented as Pearson correlation R. Cells infected with MOI = 3 at 8 and 12 hpi; data pooled from two independent experiments ( n = 6, biological replicates, **** p < 0.0001). For 24 and 48 hpi, MOI = 1 was used ( n = 3) biological replicates. Statistical analysis was performed using one-way ANOVA. Data shown as mean ± s.e.m. .

Article Snippet: The following antibodies were used for this study: anti-SARS-CoV-2 nucleocapsid mouse antibody (Cell Signaling Technology, 33717—1:200 dilution), anti-double stranded RNA J2 mouse antibody (Exalpha, 10010500—1:200 dilution), anti-calnexin rabbit antibody (Cell Signaling Technology, 2679—1:200 dilution), anti-FABP4 rabbit monoclonal antibody (Abcam, ab216708—1:200 dilution), anti-FABP4 goat polyclonal antibody (Novus, AF1443—1:200 dilution), anti-mouse secondary antibody (Cell Signaling Technology, 4410—1:1000 dilution), anti-rabbit secondary antibody (ThermoFisher Scientific, A-11037—1:1000 dilution).

Techniques: Infection, Expressing, Plaque Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Fluorescence, Staining, Control

Journal: EMBO Molecular Medicine

Article Title: FABP4 as a therapeutic host target controlling SARS-CoV-2 infection

doi: 10.1038/s44321-024-00188-x

Figure Lengend Snippet: ( A – D ) Pre-adipocytes and differentiated adipocytes infected with SARS-CoV-2 (WA1/2020, MOI = 0.1). ( A , B ) Relative expression of viral ( A ) genomic RNA (nucleocapsid, **** p < 0.0001, *** p = 0.0003 and 0.0004) and ( B ) sub-genomic RNA (ORF1ab, **** p < 0.0001), normalized to β-actin. ( C ) Viral loads measured from supernatant using plaque assay (**** p < 0.0001,*** p = 0.0001). Data are pooled from two independent experiments ( n = 8, biological replicates). Statistical analysis was performed using two-way ANOVA. ( D ) IL-6 measured by ELISA in the supernatant of differentiated adipocytes with or without virus infection (MOI = 0.1). Data are pooled from two independent experiments ( n = 8, biological replicates, **** p < 0.0001). Statistical analysis was performed using one-way ANOVA. ( E ) Western blots of SARS-CoV-2 nucleocapsid, FABP4, β-actin protein levels, and total protein (Ponceau S staining) in cell lysates of differentiated adipocytes infected with SARS-CoV-2 (MOI = 0.1 or MOI = 1). ( F ) Quantification of FABP4 band intensity normalized to total protein, representative of two independent experiments ( n = 4, biological replicates). ( G ) FABP4 gene expression relative to β-actin, pooled from two independent experiments ( n = 8, biological replicates). ( H ) FABP4 secretion in the supernatant within 1-hour incubation at the indicated time points following infection. Fold change is calculated relative to uninfected samples. Data are representative of two independent experiments ( n = 6, biological replicates, * p = 0.0336). Statistical analysis was performed using two-way ANOVA. ( I , J ) Representative confocal images of infected adipocytes stained with nucleocapsid (red), FABP4 (green), and lipid droplets (blue). ( I ) Low magnification and ( J ) high magnification images of the same samples (Scale bars = 50 μm, magnified regions = 10 μm) ( n = 3, biological replicates). ( K ) Percentage lipid droplet area relative to nucleocapsid-positive area per cell in infected differentiated adipocytes. Pearson correlation coefficient is indicated as r. Data are shown as the mean ± s.e.m. .

Article Snippet: The following antibodies were used for this study: anti-SARS-CoV-2 nucleocapsid mouse antibody (Cell Signaling Technology, 33717—1:200 dilution), anti-double stranded RNA J2 mouse antibody (Exalpha, 10010500—1:200 dilution), anti-calnexin rabbit antibody (Cell Signaling Technology, 2679—1:200 dilution), anti-FABP4 rabbit monoclonal antibody (Abcam, ab216708—1:200 dilution), anti-FABP4 goat polyclonal antibody (Novus, AF1443—1:200 dilution), anti-mouse secondary antibody (Cell Signaling Technology, 4410—1:1000 dilution), anti-rabbit secondary antibody (ThermoFisher Scientific, A-11037—1:1000 dilution).

Techniques: Infection, Expressing, Plaque Assay, Enzyme-linked Immunosorbent Assay, Virus, Western Blot, Staining, Gene Expression, Incubation

Journal: EMBO Molecular Medicine

Article Title: FABP4 as a therapeutic host target controlling SARS-CoV-2 infection

doi: 10.1038/s44321-024-00188-x

Figure Lengend Snippet: ( A ) Percentage of FABP4 bound with fatty acid (BODIPY FL C12) in the presence or absence of CRE-14 or BMS309403 ( n = 3, technical replicates, **** p < 0.0001). ( B ) Representative MST time traces with blue and red regions indicating F cold and F hot , respectively, from which fluorescence measurements were normalized. ( C ) Dose-response curve showing FABP4 binding to increasing concentrations of CRE-14, represented as normalized fluorescence. KD value (954 nM) represents the average across two technical runs. ( D ) MRC5 cell viability following administration of titrated doses of CRE-14 at the indicated concentrations of FBS. ( E , F ) Differentiated adipocytes infected with SARS-CoV-2 (WA1/2020, MOI = 0.1) and treated with either CRE-14 (20 μM) or DMSO. ( E ) Relative RNA expression of nucleocapsid normalized to β-actin (**** p < 0.0001). ( F ) Viral load measured from the supernatant using plaque assay (**** p < 0.0001, *** p = 0.0004). ( G ) Representative confocal images of control and infected adipocytes (MOI = 1), fixed 48 h post-infection, stained for virus nucleocapsid (red), lipid droplets (yellow), and nuclei (DAPI, blue) ( n = 3, biological replicates). Scale bar = 500 μm. ( H ) Percentage of nucleocapsid-positive area per image, averaging 4–5 images per sample (**** p < 0.0001). ( I ) Representative confocal images showing lipid droplet content (yellow) in WT and FABP4-deficient adipocytes. ( J ) FABP4 -shRNA knockdown and scrambled controls infected with SARS-CoV-2 (WA1/2020, MOI = 1), with viral titers measured by plaque assay from supernatants. Data are pooled from two independent experiments (n = 8, biological replicates, **** p < 0.0001). Statistical analysis was performed using two-way ANOVA. ( K – M ) WT and FABP4-deficient differentiated adipocytes infected and treated with either DMSO, CRE-14, or BMS309403 at indicated doses, with cell lysates collected 48 h post-infection. Data are representative of two independent experiments ( n = 3). ( K ) Western blots showing nucleocapsid, FABP4, and total proteins (Ponceau S staining) in cell lysates. ( L , M ) Quantifications of nucleocapsid band intensity relative to total protein. Statistical analysis was performed using two-way ANOVA (**** p < 0.0001, * p = 0.017). ( N ) Percentage of lipid droplet area per cell in uninfected and SARS-CoV-2-infected adipocytes with or without FABP4 inhibitor treatment (20 μM, **** p < 0.0001, *** p = 0.0006). ( O , P ) Adipocytes infected at 5, 10, and 20 days post-differentiation (MOI = 1). ( O ) Western blot of nucleocapsid, FABP4, and GAPDH protein levels in cell lysates, and ( P ) viral titers in supernatant measured 48 h post-infection. Data are representative of two independent experiments ( n = 3, biological replicates, **** p < 0.0001, *** p = 0.0008, ** p = 0.0016). Statistical analysis was performed using two-way ANOVA. Data are shown as mean ± s.e.m. .

Article Snippet: The following antibodies were used for this study: anti-SARS-CoV-2 nucleocapsid mouse antibody (Cell Signaling Technology, 33717—1:200 dilution), anti-double stranded RNA J2 mouse antibody (Exalpha, 10010500—1:200 dilution), anti-calnexin rabbit antibody (Cell Signaling Technology, 2679—1:200 dilution), anti-FABP4 rabbit monoclonal antibody (Abcam, ab216708—1:200 dilution), anti-FABP4 goat polyclonal antibody (Novus, AF1443—1:200 dilution), anti-mouse secondary antibody (Cell Signaling Technology, 4410—1:1000 dilution), anti-rabbit secondary antibody (ThermoFisher Scientific, A-11037—1:1000 dilution).

Techniques: Fluorescence, Binding Assay, Infection, RNA Expression, Plaque Assay, Control, Staining, Virus, shRNA, Knockdown, Western Blot

Journal: EMBO Molecular Medicine

Article Title: FABP4 as a therapeutic host target controlling SARS-CoV-2 infection

doi: 10.1038/s44321-024-00188-x

Figure Lengend Snippet: ( A – E ) SARS-CoV-2-infected differentiated adipocytes (MOI = 1), treated with either DMSO, BMS309403 (20 μM) or CRE-14 (20 μM). ( A ) Relative RNA expression of SARS-CoV-2 nucleocapsid, normalized to β-actin. Data are pooled from two independent experiments ( n = 8, biological replicates, **** p < 0.0001, * p = 0.0229). ( B ) Quantification of nucleocapsid band intensity normalized to β-actin. Data are representative of three independent experiments ( n = 3, biological replicates, **** p < 0.0001, *** p = 0.0003, * p = 0.0182). ( C ) Western blot of nucleocapsid and β-actin protein levels in cell lysates. ( D ) Viral load measured by plaque assay, pooled from two independent experiments ( n = 8, biological replicates, **** p < 0.0001), representative of four independent experiments. ( E ) IL-6 levels in the supernatant of infected adipocytes treated with FABP4 inhibitors, measured by ELISA. Data are pooled from two independent experiments ( n = 8, biological replicates, *** p = 0.0004 and 0.0001). For ( A ), ( B ), ( D ), and ( E ), statistical analysis was performed using two-way ANOVA. ( F – H ) Infected wild-type (WT) and FABP4-deficient human adipocytes (MOI = 1). ( F ) Western blot of nucleocapsid, FABP4, and β-actin protein levels in cell lysates. ( G ) Quantification of nucleocapsid band intensity normalized to β-actin, representative of two independent experiments ( n = 3, biological replicates, ** p = 0.0015). ( H ) Viral load measured by plaque assay from supernatants collected 24 and 48 h post-infection. Data are pooled from three independent experiments ( n = 9, biological replicates, **** p < 0.0001). Statistical analysis was performed using a standard t-test. ( I – L ) Infected differentiated adipocytes (MOI = 3, biological replicates, **** p < 0.0001) fixed 48 h post-infection and stained with dsRNA (red), lipid droplets (yellow), and calnexin (blue). ( J , K ) Representative images of infected cells with ( J ) inhibitor treatment or ( K ) genetic deletion of FABP4. Scale bar = 50 μm, magnified regions = 10 μm. ( I , L ) Percentage dsRNA area and mean fluorescence intensity per cell. Statistical analysis was performed using one-way ANOVA for ( I ) and a standard t-test for ( L ) ( n = 3, biological replicates, **** p < 0.0001) biological replicates and 11 to 16 images per sample. ( M – O ) Real-time electric impedance traces and their corresponding median cell index (hours) for ( M ) and ( N ) wild-type and FABP4 knockout mouse pre-adipocytes infected with coronavirus OC43 at indicated MOIs. ( O ) MRC5 cells infected with OC43 and treated with either DMSO or CRE-14. Statistical analysis was performed using a standard t-test ( n = 3, biological replicates, **** p < 0.0001, ** p = 0.0026). Data are shown as mean ± s.e.m. .

Article Snippet: The following antibodies were used for this study: anti-SARS-CoV-2 nucleocapsid mouse antibody (Cell Signaling Technology, 33717—1:200 dilution), anti-double stranded RNA J2 mouse antibody (Exalpha, 10010500—1:200 dilution), anti-calnexin rabbit antibody (Cell Signaling Technology, 2679—1:200 dilution), anti-FABP4 rabbit monoclonal antibody (Abcam, ab216708—1:200 dilution), anti-FABP4 goat polyclonal antibody (Novus, AF1443—1:200 dilution), anti-mouse secondary antibody (Cell Signaling Technology, 4410—1:1000 dilution), anti-rabbit secondary antibody (ThermoFisher Scientific, A-11037—1:1000 dilution).

Techniques: Infection, RNA Expression, Western Blot, Plaque Assay, Enzyme-linked Immunosorbent Assay, Staining, Fluorescence, Knock-Out

Journal: EMBO Molecular Medicine

Article Title: FABP4 as a therapeutic host target controlling SARS-CoV-2 infection

doi: 10.1038/s44321-024-00188-x

Figure Lengend Snippet: Syrian hamsters were infected intranasally with SARS-CoV-2 (Ank1, 100 TCID50) and treated daily with FABP4 inhibitor (CRE-14, 15 mg/kg) or vehicle. ( A ) Percent of initial body weight over time ( B ) Lung viral titers pooled from three independent experiments ( n = 18 for infected vehicle, n = 17 for CRE-14 treated, and n = 6 for each uninfected group). Statistical analysis was performed using two-way ANOVA for ( A ) (**** p < 0.0001) and a standard t-test for ( B ) (* p = 0.028). ( C , E ) Representative immunofluorescence and IHC staining of SARS-CoV-2 nucleocapsid in infected hamster lungs with or without CRE-14 treatment (Scale bar = 1 mm, n = 4). ( D ) Percentage of nucleocapsid-positive cells relative to total cell count, quantified from immunofluorescence staining ( n = 4, biological replicates, ** p = 0.0086). Statistical analysis was performed using a standard t-test. ( F ) Representative H&E staining of control and infected hamster lungs with or without inhibitor treatment (Scale bars: left = 2 mm, right = 200 μm). Low magnification images are shown in (Fig. ). ( G – L ) Pathology evaluation of lung histology in arbitrary units (A.U.), based on pathology scores ( n = 6 vehicle treated, n = 5 CRE-14 treated, further details in Table ). Statistical analysis was performed using a standard t-test (( G ) * p = 0.0391, ( H ) ** p = 0.0063, ( J ) * p = 0.0158, ( K ) * p = 0.014, ( L ) ** p = 0.002 and * p = 0.019). ( J ) Bronchial damage represents combined pathology scores of bronchial epithelial cell necrosis and presence of cellular debris in bronchi. ( L ) Alveolar damage represents combined pathology scores of alveolar epithelial cell necrosis, cellular debris in alveoli, hyaline membranes, fibrin deposition, and alveolar emphysema. ( M ) Representative Masson’s trichrome staining of control and infected hamster lungs with or without CRE-14 treatment (Scale bar = 1 mm, n = 4). Low magnification images are shown in (Fig. ). Data are shown as mean ± s.e.m. .

Article Snippet: The following antibodies were used for this study: anti-SARS-CoV-2 nucleocapsid mouse antibody (Cell Signaling Technology, 33717—1:200 dilution), anti-double stranded RNA J2 mouse antibody (Exalpha, 10010500—1:200 dilution), anti-calnexin rabbit antibody (Cell Signaling Technology, 2679—1:200 dilution), anti-FABP4 rabbit monoclonal antibody (Abcam, ab216708—1:200 dilution), anti-FABP4 goat polyclonal antibody (Novus, AF1443—1:200 dilution), anti-mouse secondary antibody (Cell Signaling Technology, 4410—1:1000 dilution), anti-rabbit secondary antibody (ThermoFisher Scientific, A-11037—1:1000 dilution).

Techniques: Infection, Immunofluorescence, Immunohistochemistry, Cell Counting, Staining, Control

Journal: EMBO Molecular Medicine

Article Title: FABP4 as a therapeutic host target controlling SARS-CoV-2 infection

doi: 10.1038/s44321-024-00188-x

Figure Lengend Snippet: Reagents and tools table

Article Snippet: The following antibodies were used for this study: anti-SARS-CoV-2 nucleocapsid mouse antibody (Cell Signaling Technology, 33717—1:200 dilution), anti-double stranded RNA J2 mouse antibody (Exalpha, 10010500—1:200 dilution), anti-calnexin rabbit antibody (Cell Signaling Technology, 2679—1:200 dilution), anti-FABP4 rabbit monoclonal antibody (Abcam, ab216708—1:200 dilution), anti-FABP4 goat polyclonal antibody (Novus, AF1443—1:200 dilution), anti-mouse secondary antibody (Cell Signaling Technology, 4410—1:1000 dilution), anti-rabbit secondary antibody (ThermoFisher Scientific, A-11037—1:1000 dilution).

Techniques: Knock-Out, Sequencing, Recombinant, shRNA, Staining, Electron Microscopy, Plasmid Preparation, Software, Isolation, cDNA Synthesis, Luciferase, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Vitamin D alleviates HFD-induced hepatic fibrosis by inhibiting DNMT1 to affect the TGFβ1/Smad3 pathway

doi: 10.1016/j.isci.2024.111262

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-FABP4 , Cell Signaling Technology , Cat#50699.

Techniques: Recombinant, Methylation, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Sequencing, Negative Control, Positive Control, Software

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: Fig. 1. 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: Fig. 2. 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: Fig. 3. 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: Fig. 4. 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: Fig. 5. 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