recombinant mouse fabp3  (Sino Biological)

Journal: bioRxiv

Article Title: Fatty acid binding proteins shape the cellular response to activation of the glucocorticoid receptor

doi: 10.1101/2021.07.02.450968

Figure Lengend Snippet: GRE-transcriptional activity of GR in the absence or presence of FABP1, FABP2, FABP3, FABP4 or FABP5 in response to vehicle control or increasing concentrations of (A) hydrocortisone (B) dexamethasone or (C) prednisolone, was assessed using a reporter gene in COS-7 cells after 24 h treatment (n=3). (D) Expression of the GR in naïve COS-7 cells or following transfection with GR determined by immunoblotting. Transcriptional activity of the endogenous or transfected GR in the presence of FABP4 in response to (E) dexamethasone or (F) prednisolone (n=3). (G) FABP translocation to the nucleus in COS-7 cells transfected with GR and GFP-FABP1 or GFP-FABP4 following 24 h treatment with vehicle or an EC 80 concentration of hydrocortisone, dexamethasone or prednisolone (n=3). Data are mean ± SEM from n independent experiments, as stated. For concentration-response curves, symbols show means and error bars, S.E.M. *** p<0.001, two-way ANOVA with Sidak’s multiple comparison test. For bar graphs, bars show the mean, error bars the S.E.M. and symbols show the independent data points for each experiment. *** p<0.001, two-way ANOVA with Dunnett’s multiple comparisons test.

Article Snippet: Immunoblotting used primary antibodies recognising FABP1 (Abcam (AB76812; rabbit, 1:300), FABP2 (gift from Dr Satoshi Kaiura, Dainippon Sumi-tomo Pharma Co. Ltd., Osaka, Japan; mouse; 1:400), FABP3 (R&D Systems MAB1678; rabbit, 1:200), FABP4 (Abcam AB92501; rabbit, 1:1000), FABP5 (Abcam AB37267; rabbit, 1:200), GR (Abcam AB183127; rabbit, 1:400) or β-actin (Cell Signaling Technology 3700S; mouse, 1:5000).

Techniques: Activity Assay, Control, Expressing, Transfection, Western Blot, Translocation Assay, Concentration Assay, Comparison

Journal: bioRxiv

Article Title: Fatty acid binding proteins shape the cellular response to activation of the glucocorticoid receptor

doi: 10.1101/2021.07.02.450968

Figure Lengend Snippet:

Article Snippet: Immunoblotting used primary antibodies recognising FABP1 (Abcam (AB76812; rabbit, 1:300), FABP2 (gift from Dr Satoshi Kaiura, Dainippon Sumi-tomo Pharma Co. Ltd., Osaka, Japan; mouse; 1:400), FABP3 (R&D Systems MAB1678; rabbit, 1:200), FABP4 (Abcam AB92501; rabbit, 1:1000), FABP5 (Abcam AB37267; rabbit, 1:200), GR (Abcam AB183127; rabbit, 1:400) or β-actin (Cell Signaling Technology 3700S; mouse, 1:5000).

Techniques: Binding Assay, Purification

Journal: Biomedicines

Article Title: Fatty Acid-Binding Proteins Aggravate Cerebral Ischemia-Reperfusion Injury in Mice

doi: 10.3390/biomedicines9050529

Figure Lengend Snippet: Effects of I/R injury on FABP3, FABP5 and FABP7 expression levels in the brain. ( A ) The locations of samples. Brains were cut into four slices (2-mm thick) from the front of the cortex. The second slices (designated as con and ips slices) of the brain, including the cortex and striatum, were used for Western blot analysis and PGE 2 -content analysis in the following experiments. ( B – E ) Mice were subjected to right tMCAO for 2 h. At 6, 12, 24 and 48 h after reperfusion, the second slice of the right brain (ipsilateral) and the left brain (contralateral) areas were collected for Western blot analysis of FABPs levels. ( B ) Representative images of Western blots. Quantitative analyses of FABP3 ( C ), FABP5 ( D ) and FABP7 ( E ) expression levels in contralateral (black) and ipsilateral (blue) areas of the brains. # p < 0.05, ## p < 0.01 vs. the sham group (contralateral); * p < 0.05, ** p < 0.01 vs. the sham group (ipsilateral) (n = 6 per group). Differences were statistically analyzed using one-way analysis of variance (ANOVA) followed by Dunnett’s test.

Article Snippet: The following working dilutions were used for the indicated monoclonal antibodies, per manufacturer’s suggestions: mouse anti-FABP3 (1:1000; Hycult Biotech, HM2016, Uden, NLD), goat anti-FABP5 (1:1000; R&D Systems, AF3077, Minneapolis, MN, USA), goat anti-FABP7 (1:1000; R&D Systems, AF3166, Minneapolis, MN, USA), rabbit anti-mPGES-1 (1:200; Cayman Chemical, 160140, Ann Arbor, MI, USA) and mouse anti-β-actin (1:5000; Sigma, A5441, St Louis, MO, USA).

Techniques: Expressing, Western Blot

Journal: Biomedicines

Article Title: Fatty Acid-Binding Proteins Aggravate Cerebral Ischemia-Reperfusion Injury in Mice

doi: 10.3390/biomedicines9050529

Figure Lengend Snippet: Immunofluorescence of FABP3, FABP5 and FABP7 in the cortexes of sham and I/R mice. ( A ) Representative micro-graphs of immunofluorescence staining of the cortical penumbra region (shown in the black box area) at 12 h after reperfusion. ( B ) Double staining for FABP3 (green) and NeuN (a neuronal marker, red) expression in sham mice ( B1 ) and I/R mice ( B2 , ipsilateral). ( C , D ) Double staining for FABP5 (green) and NeuN (red; C ) or Olig2 (an oligodendrocyte marker, red; D ) in sham mice and I/R mice (ipsilateral). ( E , F ) Double staining for FABP7 (green) and GFAP (an as-trocyte marker, red; E ) or Olig2 (red; F ). Scale bar = 50 μm. The two small images on the left show immunofluorescence for FABPs and cell markers, whereas the larger image on the right is a merged image.

Article Snippet: The following working dilutions were used for the indicated monoclonal antibodies, per manufacturer’s suggestions: mouse anti-FABP3 (1:1000; Hycult Biotech, HM2016, Uden, NLD), goat anti-FABP5 (1:1000; R&D Systems, AF3077, Minneapolis, MN, USA), goat anti-FABP7 (1:1000; R&D Systems, AF3166, Minneapolis, MN, USA), rabbit anti-mPGES-1 (1:200; Cayman Chemical, 160140, Ann Arbor, MI, USA) and mouse anti-β-actin (1:5000; Sigma, A5441, St Louis, MO, USA).

Techniques: Immunofluorescence, Staining, Double Staining, Marker, Expressing

Journal: Biomedicines

Article Title: Fatty Acid-Binding Proteins Aggravate Cerebral Ischemia-Reperfusion Injury in Mice

doi: 10.3390/biomedicines9050529

Figure Lengend Snippet: Effects of MF6 administration on FABP3, FABP5 and FABP7 expression levels in I/R mice. Mice were subjected to right tMCAO for 2 h and administered 3 mg/kg MF6 at 30 min after reperfusion. At 12 h after reperfusion, the second slice of the right (ipsilateral) and left (contralateral) brain areas, including the cortex and striatum, were used for Western blot analysis of FABP levels. ( A ) Representative images of Western blots. Quantitative analyses of FABP3 ( B ), FABP5 ( C ) and FABP7 ( D ) proteins expression levels in the contralateral (black) and ipsilateral (blue) regions of the brains. ** p < 0.01 vs. the sham-treated group, administered the vehicle (contralateral/ipsilateral); # p < 0.05, ## p < 0.01 vs. the I/R-treated group, administered the vehicle (contralateral/ipsilateral) (n = 6 per group). Differences were statistically analyzed using two-way analysis of variance (ANOVA) followed by Student–Newman–Keuls test.

Article Snippet: The following working dilutions were used for the indicated monoclonal antibodies, per manufacturer’s suggestions: mouse anti-FABP3 (1:1000; Hycult Biotech, HM2016, Uden, NLD), goat anti-FABP5 (1:1000; R&D Systems, AF3077, Minneapolis, MN, USA), goat anti-FABP7 (1:1000; R&D Systems, AF3166, Minneapolis, MN, USA), rabbit anti-mPGES-1 (1:200; Cayman Chemical, 160140, Ann Arbor, MI, USA) and mouse anti-β-actin (1:5000; Sigma, A5441, St Louis, MO, USA).

Techniques: Expressing, Western Blot