rabbit polyclonal anti fabp5  (Cusabio)

Journal: iScience

Article Title: Astrocytic FABP5 mediates retrograde endocannabinoid transport at central synapses

doi: 10.1016/j.isci.2025.112342

Figure Lengend Snippet: FABP5 is essential for 2-AG-mediated DSI at CA1 GABA synapses (A) Immunolabelling of FABP5 expression in the hippocampal CA1 region of WT mice. a 1- a 2 , Labeling of FABP5 and the astrocyte marker s100β. a 3 , Triple labeling between the neuronal marker NeuN, FABP5, and s100β. Note the robust colocalization of FABP5 with s100β. Scale bar: 100 μm. a 4 , Higher magnification of NeuN, FABP5 and s100β triple labeling. Scale bar: 10 μm. (B) Lack of FABP5 immunostaining in the hippocampus of FABP5 KO mice. b 1, b 2 , FABP5 and s100β staining. b 3 , Merged image of FABP5, s100β, and DAPI labeling. Scale bar: 100 μm. b 4 , merged image of FABP5, NeuN, and DAPI labeling. Scale bar: 100 μm. (C) Inhibition of FABP5 blunts hippocampal DSI. Left panel illustrates averaged magnitude of DSI obtained in WT ( : 47.41 ± 1.11%; n = 8 cells; N = 3 mice), FABP5 KO ( : 5.26 ± 2.37%; n = 8 cells; N = 3 mice; p = 1.95x10 −11 versus WT), and following FABP5 inhibition with 10 μM SBFI-103 ( : 6.59 ± 2.72%; n = 8 cells; N = 3 mice; p = 3.59x10 −11 versus WT). Upper right panel depicts representative traces of IPSCs recorded before and during DSI. Scale bars: 100 ms, 200 pA. Lower right panel represents the time course of DSI. (D) Deletion of FABP5 does not alter the function of CB1Rs. Upper panel illustrates representative IPSCs traces collected before (1) and during the application of WIN55,212-2 (10 μM) (2) from WT ( ) and KO ( ). Lower panel is a summary of the depression of IPSC amplitude induced by WIN55,212-2 in WT ( : 42.53 ± 6.88%; n = 7 cells; N = 3 mice; p = 1.75x10 −7 versus baseline) and KO ( : 43.01 ± 7.24%; n = 7 cells; N = 3 mice; p = 1.44x10 −7 versus baseline). Scale bars: 50 ms, 50 pA. Data are represented as mean ± SEM. ∗∗∗∗ p < 0.001. one-way ANOVA with Bonferroni’s multiple comparisons test.

Article Snippet: Hippocampal sections (30 μm) were incubated with the following primary antisera: rabbit anti-FABP5 (BioVendor R&D, RD181060100), goat anti-FABP5 (R&D Systems Inc, #AF1476), mouse anti-NeuN (Millipore, MAB377), rabbit anti-NeuN (Abcam, #AB104225), and mouse anti-s100β (Sigma, #S2532).

Techniques: Expressing, Labeling, Marker, Immunostaining, Staining, Inhibition

Journal: iScience

Article Title: Astrocytic FABP5 mediates retrograde endocannabinoid transport at central synapses

doi: 10.1016/j.isci.2025.112342

Figure Lengend Snippet: Expression of FABP5 in FABP5 KO mice restores hippocampal DSI (A) AAV-mediated expression of FABP5 in astrocytes of the hippocampal CA1 region of FABP5 KO mice. a 1 , Low magnification image of FABP5, DAPI, and s100β labeling. Scale bar: 100 μm. a 2 -a 4 , High magnification labeling of FABP5, s100β, and merge. Scale bar: 10 μm. (B) AAV-mediated expression of FABP5 in neurons of FABP5 KO mice. b 1 , Low magnification labeling of DAPI, NeuN, and FABP5. Scale bar: 100 μm. b 2 -b 4 , High magnification labeling of FABP5, NeuN, and merge with DAPI. Scale bars: 10 μm. (C) Upper image illustrates GFP expression in the CA1 region. Lower image shows recording and stimulating electrodes in the same CA1 region expressing FABP5. (D) Re-expression of FABP5 restores DSI in FABP5 KO mice. D 1 , Averaged magnitude of the DSI in WT ( : 43.18 ± 3.16%; n = 10 cells; N = 3 mice), KO ( : 1.76 ± 0.76%; n = 9 cells; N = 3 mice; p = 5.96x10 −14 versus WT), KO+AAV-CAG-FABP5 ( : 39.63 ± 2.55%; n = 9 cells; N = 3 mice; p = 1 versus WT), and KO+AAV-CAG-EGFP ( : 7.15 ± 1.73%; n = 11 cells; N = 4 mice; p = 2.75x10 −11 versus KO+AAV-CAG-FABP5). D 2 , Summary graph of the time course of the DSI obtained in WT (●), KO ( ), KO+AAV-CAG-FABP5 ( ), and KO+AAV-CAG-EGFP ( ). D 3 , Representative traces of IPSCs recorded before and during DSI from WT ( ), KO ( ), KO+AAV-CAG-FABP5 ( ), and KO+AAV-CAG-EGFP ( ). Scale bars: 50 ms, 200 pA. Data are represented as mean ± SEM. ∗∗∗ p < 0.001, one-way ANOVA, with Bonferroni’s multiple comparisons test.

Article Snippet: Hippocampal sections (30 μm) were incubated with the following primary antisera: rabbit anti-FABP5 (BioVendor R&D, RD181060100), goat anti-FABP5 (R&D Systems Inc, #AF1476), mouse anti-NeuN (Millipore, MAB377), rabbit anti-NeuN (Abcam, #AB104225), and mouse anti-s100β (Sigma, #S2532).

Techniques: Expressing, Labeling

Journal: iScience

Article Title: Astrocytic FABP5 mediates retrograde endocannabinoid transport at central synapses

doi: 10.1016/j.isci.2025.112342

Figure Lengend Snippet: FABP5 MUT fails to rescue hippocampal DSI in FABP5 KO mice (A) Astrocytic expression of FABP5 MUT in the CA1 region of FABP5 KO mice. a 1 , Low magnification image of FABP5, s100β, and DAPI labeling in the CA1 region. Scale bar: 100 μm. a 2 -a 4 , High magnification immunostaining of FABP5 labeling, s100β, and merge. Scale bar: 10 μm. (B) Neuronal expression of FABP5 MUT in the CA1 region of FABP5 KO mice. b 1 , Low magnification image of FABP5, NeuN, and DAPI labeling in the CA1 region. Scale bar: 100 μm. b 2 -b 4 , High magnification immunostaining of FABP5, NeuN, and merge with DAPI. Scale bar: 10 μm. (C) Expression of FABP5 MUT in FABP5 KO mice did not rescue hippocampal DSI. Left panel, summary of time course of DSI recorded from wild-type WT+AAV-CAG-FABP5 MUT ( ), WT+AAV-CAG-EGFP ( ), KO+AAV-CAG-FABP5 MUT ( ), and KO+AAV-CAG-EGFP ( ). Right panel, Sample superimposed IPSC traces collected before and during the DSI. Scale bars: 50 ms, 200 pA (D) Summary of DSI magnitude obtained in WT+AAV-CAG-FABP5 MUT ( : 45.86 ± 5.67%; n = 10 cells; N = 3 mice), WT+AAV-CAG-EGFP ( : 44.28 ± 2.86%; n = 11 cells; N = 3 mice), KO+AAV-CAG-FABP5 MUT ( : 3.19 ± 1.27%; n = 13 cells; N = 3 mice; p = 1.54×10 −11 versus WT+AAV-CAG-FABP5 MUT ), KO+AAV-CAG-EGFP ( : 7.15 ± 1.73%; n = 11 cells; N = 3 mice; p = 1 versus KO+AAV-CAG-FABP5 MUT , p = 1.05×10 −9 versus WT+AAV-CAG-EGFP. (E) Expression of FABP5 MUT in FABP5 KO mice does not affect the depression of IPSC amplitude induced by WIN55,212-2 (10 μM) ( , 34.75 ± 68.28%; n = 7; p = 2.21×10 −4 versus baseline). Inset, Sample IPSCs traces collected before and during the application of WIN55,212-2 (10μM). Scale bars: 25 ms, 50 pA. Data are represented as mean ± SEM. ∗∗∗ p < 0.001, one-way ANOVA with Bonferroni’s multiple comparisons test, or paired sample t-test.

Article Snippet: Hippocampal sections (30 μm) were incubated with the following primary antisera: rabbit anti-FABP5 (BioVendor R&D, RD181060100), goat anti-FABP5 (R&D Systems Inc, #AF1476), mouse anti-NeuN (Millipore, MAB377), rabbit anti-NeuN (Abcam, #AB104225), and mouse anti-s100β (Sigma, #S2532).

Techniques: Expressing, Labeling, Immunostaining

Journal: iScience

Article Title: Astrocytic FABP5 mediates retrograde endocannabinoid transport at central synapses

doi: 10.1016/j.isci.2025.112342

Figure Lengend Snippet: Extracellular pool of FABP5 mediates hippocampal DSI (A) Secreted FABP5 variant (FABP5 SEC ) binds to 2-AG with comparable affinity to WT FABP5 (FABP5 Ki: 1.0 ± 0.1 μM; FABP5 SEC Ki: 1.6 ± 0.2 μM, n = 4). (B) AAV-mediated expression of FABP5 SEC in CA1 astrocytes of FABP5 KO mice. b 1 -b 3 , Immunolabelling for FABP5, s100β, and merge with DAPI. Scale bar: 10 μm. (C) Neuronal expression of FABP5 SEC in the CA1 region. c 1 -c 3 , Immunolabelling for FABP5, NeuN, and merge with DAPI. Scale bar: 10 μm. (D) Expression of FABP5 SEC but not FABP7 restores DSI in FABP5 KO mice. D 1 , left panel, Summary of the time course of hippocampal DSI recorded from WT ( ), KO ( ), KO+AAV-CAG-EGFP ( ), KO+AAV-CAG-FABP5 SEC ( ), and KO+AAV-CAG-FABP7 ( ). Right panel, Superimposed IPSC traces collected before and during DSI. D 2 , Averaged magnitude of DSI obtained in WT ( : 45.99 ± 2.51%; n = 10 cells; N = 5 mice), KO ( : 5.85 ± 1.21%; n = 17 cells; N = 5 mice), KO+AAV-CAG-EGFP ( : 10.24 ± 1.70%; n = 16 cells; N = 5 mice), KO+AAV-CAG-FABP5 SEC ( : 38.61 ± 1.94%; n = 17 cells; N = 5 mice; p = 9.83x10 −16 versus KO+AAV-CAG-EGFP, p = 0.16 vs. WT), and KO+AAV-CAG-FABP7 ( : 8.10 ± 2.19%; n = 18 cells; N = 5 mice; p = 3.62x10 −19 versus WT, p = 1 versus KO+AAV-CAG-EGFP, p = 8.22x10 −18 versus KO+AAV-CAG-FABP5 SEC ). Scale bars: 100 ms, 200 pA (E and F) Expression of FABP7 in the CA1 region of FABP5 KO mice. e 1 -e 3 , Immunolabelling for FABP7, s100β and merge. Scale bar: 10 μm. f 1 -f 3 , immunolabelling for FABP7, NeuN and merge. Scale bar: 10 μm. Data are represented as mean ± SEM. ∗∗∗ p < 0.001, one-way ANOVA with Bonferroni’s multiple comparisons test.

Article Snippet: Hippocampal sections (30 μm) were incubated with the following primary antisera: rabbit anti-FABP5 (BioVendor R&D, RD181060100), goat anti-FABP5 (R&D Systems Inc, #AF1476), mouse anti-NeuN (Millipore, MAB377), rabbit anti-NeuN (Abcam, #AB104225), and mouse anti-s100β (Sigma, #S2532).

Techniques: Variant Assay, Expressing

Journal: iScience

Article Title: Astrocytic FABP5 mediates retrograde endocannabinoid transport at central synapses

doi: 10.1016/j.isci.2025.112342

Figure Lengend Snippet: Conditional deletion of hippocampal FABP5 blunts DSI (A) Schematic of the gene structure of FABP5 FLOX mice. (B) Expression of FABP5 and a lack of tdTomato in the hippocampus of FABP5 FLOX mice. Left panel, Immunostaining for DAPI and tdTomato in CA1 region of FABP5 FLOX mice. Right panel, Immunostaining for DAPI and FABP5 in CA1 region of FABP5 FLOX mice. Scale bar: 50 μm. (C) Cre-mediated deletion of FABP5 and expression of tdTomato in the CA1 region of FABP5 FLOX mice. c1-c3, Immunostaining for tdTomato, s100β, and merge. c4-c6, Immunolabeling of tdTomato, NeuN and merge. c7-c9, immunolabeling of tdTomato, FABP5 and merge. Scale bar: 50 μm. (D) Conditional deletion of FABP5 inhibits DSI. Left panel, Averaged magnitude of hippocampal DSI recorded in FABP5 FLOX ( : 41.29 ± 3.54%; n = 11 cells; N = 5 mice), FABP5 FLOX +AAV-CMV-Cre ( : 3.51 ± 1.22%; n = 11 cells; N = 3 mice; p = 3.95×10 −12 versus FABP5 FLOX ), FABP5 FLOX +AAV-CMV-EGFP ( : 34.61 ± 2.93%; n = 11 cells; N = 4 mice; p = 1.44×10 − 9 versus FABP5 FLOX +AAV-CMV-Cre, p = 0.87 versus FABP5 FLOX ). Right panel, representative traces of IPSCs collected before and during DSI from FABP5 FLOX ( ), FABP5 FLOX +AAV-CMV-Cre ( ), and FABP5 FLOX +AAV-CMV-EGFP ( ). Scale bars: 100 ms, 200 pA (E) Averaged time course of the DSI recorded in FABP5 FLOX and FABP5 FLOX +AAV-CMV-EGFP mice. (F) Conditional deletion of FABP5 in the CA1 region does not affect the function of presynaptic CBRs. Left panel, Summary of the depression of IPSC amplitude induced by WIN55,212-2 (10 μM) in FABP5 FLOX ( : 46.13 ± 9.15%; n = 7 cells; N = 4 mice; p = 7.03×10 −5 versus baseline) and FABP5 FLOX +AAV-CMV-Cre ( : 38.76 ± 9.63%; n = 6 cells; N = 4 mice; p = 2.55×10 −5 versus baseline). Right panel, superimposed IPSCs traces collected before and during WIN55,212-2 application. Scale bars: 100 ms, 200 pA. Data are represented as mean ± SEM. ∗∗∗ p < 0.001, one-way ANOVA with Bonferroni’s multiple comparisons test.

Article Snippet: Hippocampal sections (30 μm) were incubated with the following primary antisera: rabbit anti-FABP5 (BioVendor R&D, RD181060100), goat anti-FABP5 (R&D Systems Inc, #AF1476), mouse anti-NeuN (Millipore, MAB377), rabbit anti-NeuN (Abcam, #AB104225), and mouse anti-s100β (Sigma, #S2532).

Techniques: Expressing, Immunostaining, Immunolabeling

Journal: iScience

Article Title: Astrocytic FABP5 mediates retrograde endocannabinoid transport at central synapses

doi: 10.1016/j.isci.2025.112342

Figure Lengend Snippet: Deletion of astrocytic FABP5 impairs hippocampal DSI (A) Schematic showing the experimental strategy used to delete astrocytic FABP5 and generate FABP5 FLOX /Aldh1l1 Cre mice. FABP5 FLOX /Aldh1l1 Cre /Tam represent tamoxifen-injected FABP5 FLOX /Aldh1l1 Cre mice, FABP5 FLOX /Aldh1l1/Tam represent tamoxifen-injected Cre − littermates, while FABP5 FLOX /Aldh1l1 Cre mice received vehicle. (B) Tamoxifen-induced deletion of the FABP5 and expression of tdTomato in astrocytes in the CA1 region of FABP5 FLOX /Aldh1l1 Cre /Tam mice. b1-b3, Images of tdTomato, NeuN staining, and merge. b4-b6, Images of tdTomato, s100β, and merge. b7-b9, images of immunostaining for tdTomato, FABP5, and merge. Note the expression of tdTomato in astrocytes but not in neurons and the absence of FABP5 expression. Scale bars: 50 μm. (C) Tamoxifen-induced deletion of FABP5 impairs DSI in FABP5 FLOX /Aldh1l1 Cre /Tam mice. Left panel, Averaged DSI magnitude obtained in FABP5 FLOX /Aldh1l1 Cre ( : 38.12 ± 2.97%; n = 16 cells; N = 4 mice), in FABP5 FLOX/ Aldh1l1/Tam ( : 32.12 ± 2.50%; n = 15 cells; N = 4 mice; p = 0.27 versus FABP5 FLOX /Aldh1l1 Cre ) and in FABP5 FLOX /Aldh1l1 Cre /Tam ( : 6.32 ± 1.85%; n = 14 cells; N = 4 mice; p = 2.22×10 −8 versus FABP5 FLOX /Aldh1l1/Tam). Right panel, Superimposed IPSC traces collected before and during DSI from FABP5 FLOX /Aldh1l1 Cre ( ), FABP5 FLOX /Aldh1l1/Tam ( ), and FABP5 FLOX /Aldh1l1 Cre /Tam ( ) mice. Scale bars: 100 ms, 200 pA. (D) Averaged time course of DSI recorded from FABP5 FLOX /Aldh1l1 Cre ( ), FABP5 FLOX /Aldh1l1/Tam ( ), and FABP5 FLOX /Aldh1l1 Cre /Tam ( ) mice. (E) Deletion of astrocytic FABP5 does not affect the function of CB1Rs. Summary of the depression of IPSC amplitude induced by WIN55,212-2 (10 μM) in FABP5 FLOX /Aldh1l1 Cre ( :44.75 ± 3.73%; n = 5 cells; N = 3 mice; p = 2.52×10 − 6 versus baseline) and FABP5 FLOX /Aldh1l1 Cre /Tam ( : 54.74 ± 8.11%; n = 5 cells; N = 3 mice; p = 2.29×10 − 5 versus baseline). Right panel, Sample IPSCs traces collected before and during WIN55,212-2 application. Scale bars: 100 ms, 100 pA. Data are represented as mean ± SEM. ∗∗∗ p < 0.001, one-way ANOVA with Bonferroni’s multiple comparisons test.

Article Snippet: Hippocampal sections (30 μm) were incubated with the following primary antisera: rabbit anti-FABP5 (BioVendor R&D, RD181060100), goat anti-FABP5 (R&D Systems Inc, #AF1476), mouse anti-NeuN (Millipore, MAB377), rabbit anti-NeuN (Abcam, #AB104225), and mouse anti-s100β (Sigma, #S2532).

Techniques: Injection, Expressing, Staining, Immunostaining

Journal: iScience

Article Title: Astrocytic FABP5 mediates retrograde endocannabinoid transport at central synapses

doi: 10.1016/j.isci.2025.112342

Figure Lengend Snippet: Expression of FABP5 in astrocytes of FABP5 KO mice rescues DSI (A) AAV-mediated expression of FABP5 under the control of the gfaABC1D promoter in FABP5 KO mice. a1-a3, Immunostaining of FABP5, s100β and merge. Scale bar: 50 μm. a4, Co-localization of FABP5 with s100β. a5, Lack of co-localization of FABP5 with NeuN. Scale bars: 10 μm. (B) Expression of FABP5 in astrocytes rescues DSI in FABP5 KO mice. Left panel, averaged magnitude of DSI obtained in WT ( : 42.23 ± 3.44%; n = 12 cells; N = 3 mice), KO ( : 2.65 ± 1.03%; n = 13 cells; N = 3 mice; p = 3.28×10 −13 versus WT), KO+AAV-gfaABC1D-FABP5 ( : 33.37 ± 3.50%; n = 13 cells; N = 4 mice; p = 0.12 versus WT), and KO+AAV-GFAP-EGFP ( : 6.07 ± 1.58%; n = 12 cells; N = 4 mice; p = 1.71×10 − 8 versus KO+AAV-gfaABC1D-FABP5). Right panel, IPSC traces collected before and during DSI from WT ( ), KO ( ), KO+AAV-gfaABC1D-FABP5 ( ), and KO+AAV-GFAP-EGFP ( ). Scale bars: 100 ms, 200 pA. (C) AAV-mediated expression of FABP5 under the neuron-specific hsynapsin1 promoter in FABP5 KO mice. c1-c3, Immunostaining for NeuN, FABP5, and merge. Scale bar: 50 μm. c4, Co-localization of FABP5 with NeuN. c5, Lack of co-localization of FABP5 with s100β. Scale bars: 10 μm. (D) Expression of FABP5 in neurons partially rescues DSI in FABP5 KO mice. Left panel, Summary of DSI magnitude recorded from WT ( : 39.52 ± 1.65%; n = 11 cells; N = 5 mice), KO ( : 1.43 ± 0.66%; n = 11 cells; N = 4 mice; p = 4.67×10 −18 versus WT), KO+AAV-hSyn-FABP5 ( : 13.95 ± 2.13%; n = 15 cells; N = 5 mice; p = 4.47×10 −13 versus WT) and KO+AAV-hSyn-EGFP ( : 5.15 ± 1.61%; n = 12 cells; N = 4 mice; p = 0.89 versus KO). Right panel, IPSC traces collected before and during DSI. Data are represented as mean ± SEM. ∗ p < 0.05; ∗∗∗ p < 0.001, one-way ANOVA with Bonferroni’s multiple comparisons test.

Article Snippet: Hippocampal sections (30 μm) were incubated with the following primary antisera: rabbit anti-FABP5 (BioVendor R&D, RD181060100), goat anti-FABP5 (R&D Systems Inc, #AF1476), mouse anti-NeuN (Millipore, MAB377), rabbit anti-NeuN (Abcam, #AB104225), and mouse anti-s100β (Sigma, #S2532).

Techniques: Expressing, Control, Immunostaining

Journal: iScience

Article Title: Astrocytic FABP5 mediates retrograde endocannabinoid transport at central synapses

doi: 10.1016/j.isci.2025.112342

Figure Lengend Snippet:

Article Snippet: Hippocampal sections (30 μm) were incubated with the following primary antisera: rabbit anti-FABP5 (BioVendor R&D, RD181060100), goat anti-FABP5 (R&D Systems Inc, #AF1476), mouse anti-NeuN (Millipore, MAB377), rabbit anti-NeuN (Abcam, #AB104225), and mouse anti-s100β (Sigma, #S2532).

Techniques: Virus, Recombinant, Software, Imaging, Transfection, Real-time Polymerase Chain Reaction, SYBR Green Assay, Mass Spectrometry, Control, Microscopy

Journal: Journal of Clinical Medicine

Article Title: Decreased PAX6 and DSG1 Protein Expression in Corneal Epithelium of Patients with Epithelial Basal Membrane Dystrophy, Salzmann Nodular Degeneration, and Pterygium

doi: 10.3390/jcm14051456

Figure Lengend Snippet: Primers used for RT-qPCR of mRNAs and miRNAs.

Article Snippet: FABP5 rabbit polyclonal , 1:1000 , 12348-1-AP , Proteintech, Planegg-Martinsried, Germany.

Techniques: Binding Assay

Journal: Journal of Clinical Medicine

Article Title: Decreased PAX6 and DSG1 Protein Expression in Corneal Epithelium of Patients with Epithelial Basal Membrane Dystrophy, Salzmann Nodular Degeneration, and Pterygium

doi: 10.3390/jcm14051456

Figure Lengend Snippet: Primary antibodies used for Western blot.

Article Snippet: FABP5 rabbit polyclonal , 1:1000 , 12348-1-AP , Proteintech, Planegg-Martinsried, Germany.

Techniques: Western Blot

Journal: Journal of Clinical Medicine

Article Title: Decreased PAX6 and DSG1 Protein Expression in Corneal Epithelium of Patients with Epithelial Basal Membrane Dystrophy, Salzmann Nodular Degeneration, and Pterygium

doi: 10.3390/jcm14051456

Figure Lengend Snippet: ADH7, ALDH1A1, and FABP5 mRNA expression in conjunctival impression cytology (IC) ( A , D , G ) and corneal epithelial samples ( B , E , H ) and ADH7, ALDH1A1, and FABP5 protein expression in corneal epithelial samples ( C , F , I ) of EBMD, SND, pterygium, congenital aniridia, and healthy control subjects (ctrl). Relative levels of mRNA expression were determined by RT-qPCR and protein expression by Western blot analysis. Expression of target genes is presented as geometric mean (RT-qPCR) and arithmetic mean (protein expression) with corresponding standard deviations and a representative Western blot picture. Significances are indicated (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: FABP5 rabbit polyclonal , 1:1000 , 12348-1-AP , Proteintech, Planegg-Martinsried, Germany.

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot

Journal: Journal of Clinical Medicine

Article Title: Decreased PAX6 and DSG1 Protein Expression in Corneal Epithelium of Patients with Epithelial Basal Membrane Dystrophy, Salzmann Nodular Degeneration, and Pterygium

doi: 10.3390/jcm14051456

Figure Lengend Snippet: Overview of gene and protein expression in epithelial basal membrane dystrophy, Salzmann’s nodular degeneration, and pterygium.

Article Snippet: FABP5 rabbit polyclonal , 1:1000 , 12348-1-AP , Proteintech, Planegg-Martinsried, Germany.

Techniques: Expressing, Membrane

Journal: Journal of translational medicine

Article Title: Helicobacter pylori infection exacerbates metabolic dysfunction-associated steatotic liver disease through lipid metabolic pathways: a transcriptomic study.

doi: 10.1186/s12967-024-05506-y

Figure Lengend Snippet: Fig. 7 qRT-PCR and IHC validated DEGs expression in mouse liver. Critical lipid metabolism factor Srebf1(A), Fgf21 (B) inflammatory factor Il-1β (C), Tnf-α (D). DEGs Fabp5, Ppar-γ were expressed in mouse liver (E, F). Results of Fabp5 IHC staining in mouse liver in each group (G) and quantification bar graph (H). 200× magnification under a light microscope. Data are expressed as mean ± SD, n = 6, *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: The slices were treated with 3% H2O2 at room temperature for 20 min, followed by incubation with goat serum for 20 min, and subsequently with anti-FABP5 rabbit polyclonal antibody (Proteintech, Wuhan;1:100) overnight at 4 °C.

Techniques: Quantitative RT-PCR, Expressing, Immunohistochemistry, Light Microscopy

Journal: Cell death & disease

Article Title: Fatty acid-binding protein 5 is a functional biomarker and indicator of ferroptosis in cerebral hypoxia.

doi: 10.1038/s41419-024-06681-y

Figure Lengend Snippet: Fig. 2 FABP5 is concentrated at cell surface during ferroptosis. A Immunohistochemistry staining in normal human cerebral cortex sections showing endogenous expression using antibodies against CALML5, FABP5, CTSV, LGALS7, S100A14 as well as TfR1 identified previously in [28]). The data shown here are derived from the Human Protein Atlas [68]. B Normalized fluorescence intensity heat map of candidate biomarker proteins following treatment with equivalent death-inducing concentrations of different pharmacological agents in HT-1080 cells. C Validation of FABP5 antibody specificity. FABP5 overexpressing HT-1080 cells (FABP5 OE) with C-terminal FLAG tag compared to control (Ctrl) by fluorescence staining and Western blot. Results observed with two antibodies (see Supplemental Reagents Table) were indistinguishable. Right panel, FABP5 expression level was detected by whole membrane Western blot in given populations. D Timecourse of lipid peroxidation in HT-1080 as indicated by flow cytometry analysis of Bodipy C-11 fluorescence and FABP5 cell surface staining in unpermeabilized cells and confocal images of the same cells.

Article Snippet: Cells were blocked in 5% BSA Fraction V, 10% fetal bovine serum, 0.3% Triton-X in PBS for one hour then incubated in primary FABP5 antibody (Cusabio) overnight at 4 oC followed by two times PBS washes and further incubation with the secondary antibody for one hour.

Techniques: Immunohistochemistry, Staining, Expressing, Derivative Assay, Biomarker Discovery, FLAG-tag, Control, Western Blot, Membrane, Cytometry

Journal: Cell death & disease

Article Title: Fatty acid-binding protein 5 is a functional biomarker and indicator of ferroptosis in cerebral hypoxia.

doi: 10.1038/s41419-024-06681-y

Figure Lengend Snippet: Fig. 3 FABP5 is specifically upregulated during ferroptosis. A Timecourse in hours (h) of RSL3 (200 nM) or staurosporine (50 nM) induced changes in FABP5 as detected by confocal microscopy in FABP5 OE cells. B High-content analysis of FABP5 in HT-1080 cells is shown as normalized mean fluorescent intensity ± SD of n = 4 replicate samples representative of at least three independent repetitions of the experiment in A treated with RSL3 (200 nM) or staurosporine (50 nM). Western blots show FABP5 protein detection at the same time points following treatment. C Relative FABP5 expression by quantitative PCR with RSL3 (200 nM) time course. Values are shown as mean ± SD of n = 3 technical replicates related to untreated (0 h). Statistics were calculated using two-way ANOVA against respective control conditions.

Article Snippet: Cells were blocked in 5% BSA Fraction V, 10% fetal bovine serum, 0.3% Triton-X in PBS for one hour then incubated in primary FABP5 antibody (Cusabio) overnight at 4 oC followed by two times PBS washes and further incubation with the secondary antibody for one hour.

Techniques: Confocal Microscopy, High Content Screening, Western Blot, Expressing, Real-time Polymerase Chain Reaction, Control

Journal: Cell death & disease

Article Title: Fatty acid-binding protein 5 is a functional biomarker and indicator of ferroptosis in cerebral hypoxia.

doi: 10.1038/s41419-024-06681-y

Figure Lengend Snippet: Fig. 4 FABP5 biomarker validation in RSL3-treated and GPX4 knockout cells. A FABP5 fluorescence intensity changes after RSL3 or GPX4 knockout (KO) in cell lines of different etiologies. Calu-1, HCC827, HT-1080 and U-138MG cells after treatment with RSL3 (200 nM, 5 h), and SH- SY5Y and human fetal fibroblast (hFF) cells with RSL3 (200 nM) + ammonium ferric citrate (FAC, 250 μM) for 3 h. 2 × 103 cells were assayed per condition. RSL3 final intensity is shown as mean ± SD of n = 3 independent experiments with three replicate wells each. Fold change at 96 h post infection with GPX4 KO or control (empty vector) lentivirus is shown as mean ± SD of at least three independent experiments with 2 × 103 cells assayed per condition in each of three replicate wells. SH-SY5Y and hFF are viable following GPX4 KO-induced lentivirus. B Live cell brightfield and fluorescence images of HT-1080 viability (indicated by DAPI penetration) were taken hours post infection (p.i.) with GPX4 KO and control virus at 72 and 96 h. Increased loss of viability was observed at 96 h, while early detection of FABP5 antigen at 72 h is shown in C when only a fraction of cells have died. C Western analysis of respective proteins in HT-1080 cells 72 h following GPX4 ablation. Statistics were calculated using two-way ANOVA (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns not significant).

Article Snippet: Cells were blocked in 5% BSA Fraction V, 10% fetal bovine serum, 0.3% Triton-X in PBS for one hour then incubated in primary FABP5 antibody (Cusabio) overnight at 4 oC followed by two times PBS washes and further incubation with the secondary antibody for one hour.

Techniques: Biomarker Discovery, Knock-Out, Infection, Control, Plasmid Preparation, Virus, Western Blot

Journal: Cell death & disease

Article Title: Fatty acid-binding protein 5 is a functional biomarker and indicator of ferroptosis in cerebral hypoxia.

doi: 10.1038/s41419-024-06681-y

Figure Lengend Snippet: Fig. 5 Elevated FABP5 expression induces PUFA-containing lipids and ferroptosis sensitivity. A Viability of FABP5 OE compared to control HT-1080 cells treated with ferroptosis inducer IKE (1.25 µM, 16 h, two-way ANOVA (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns not significant)). B Viability of FABP5 knockdown (KD) compared to control HT-1080 cells treated with ferroptosis inducer IKE (1.25 µM, 16 h, two- way ANOVA (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns not significant)). C, D Total lipid abundance and distribution by peak values as revealed by mass spectrometry for FABP5 OE cells compared to control cells. No significant differences were observed in total lipid content. Class terms are derived from LIPIDMAPS [69]. E Lipid heatmap annotations showing z-scores of all significantly misregulated phospholipid species in FABP5 OE and control cell samples compared with controls. Nearly all misregulated lipids are classified as PUFA-containing glycerophospholipids (boxed entries) with the number of double bonds indicated after the colon. DG diacylglycerol, LPA lysophosphatidic acid, LPE 1-acyl-sn-glycero-3-phosphoethanolamine, LPS 1-acyl-sn-glycero-3-phosphoserine, PC phosphatidylcholine, PE phosphatidyletha- nolamine, PE phosphatidylinositol, PS phosphatidylserine, O ether lipids (Welch t-test, n = 3, ns not significant). F Cumulative changes in each saturation class of phospholipids up- or downregulated in FABP5 OE cells. Lipids downregulated represent a smaller fraction of the total lipid pool, as represented by the proportion of the ‘down’ pie chart. Volcano plots demonstrate fold changes of individual lipids according to saturation, the dotted line indicates significant lipids shown in (E). MUFA monounsaturated, SFA saturated, PUFA polyunsaturated.

Article Snippet: Cells were blocked in 5% BSA Fraction V, 10% fetal bovine serum, 0.3% Triton-X in PBS for one hour then incubated in primary FABP5 antibody (Cusabio) overnight at 4 oC followed by two times PBS washes and further incubation with the secondary antibody for one hour.

Techniques: Expressing, Control, Knockdown, Mass Spectrometry, Derivative Assay