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: bioRxiv

Article Title: Lysosomes cell autonomously regulate myeloid cell states and immune responses

doi: 10.1101/2024.11.11.623074

Figure Lengend Snippet: Grn KO myeloid cells display an MG8-like signature in vitro . (a) Average expression of top 150 regulated genes [from Grn KO primary microglia (pMG) in vitro ] in each microglia subcluster from aged Grn KO microglia scRNA-seq. (b) Heatmap of relative expression of top 175 MG8 markers (from aged Grn KO microglia scRNA-seq) in WT and Grn KO pMG in vitro . (c) Volcano plot displaying differential expression between Grn KO and WT pMG. Top MG8 marker genes are highlighted as triangles. (d,e) Western blots and quantification of MG8 markers (GPNMB, FABP5, and total CatB) and LPL (not expressed by MG8) from protein lysates of WT and Grn KO mouse primary microglia (pMG) (n=5 biological replicates/genotype). (f) Average expression of top 150 upregulated genes [from Grn KO bone marrow-derived macrophages (BMDMs) in vitro ] in each microglia subcluster from aged Grn KO microglia scRNA-seq. (g) Heatmap of relative expression of top 175 MG8 markers (from aged Grn KO microglia scRNA-seq) in WT and Grn KO BMDMs in vitro . (h) Volcano plot displaying differential expression between Grn KO and WT BMDMs. Top MG8 marker genes are highlighted as triangles. (i) Western blots and quantification of MG8 markers (GPNMB, FABP5, and total CatB) and LPL (not expressed by MG8) from protein lysates of WT and Grn KO mouse BMDMs (n=3 biological replicates/genotype). Data presented in (e,j) are mean±s.e.m. Student’s t-tests were performed to compare across genotypes in (e,j). * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: The following primary antibodies were used: sheep anti-PGRN (R&D Systems; Cat# 2557; 0.5μg/mL), rabbit anti-GPNMB (Cell Signaling Technology; Cat# 90205; 1:500), rabbit anti-FABP5 (Cell Signaling Technology; Cat# 39926; 1:500), rabbit anti-CatB (Cell Signaling Technology; Cat# 31718; 1:500), rabbit anti-LPL (Biotechne; Cat# AF7197; 1μg/mL), mouse anti-Vinculin (Sigma-Aldrich; Cat# V9264; 1:1000), mouse anti-β-Actin (Sigma-Aldrich; Cat# A2228; 1:10000), mouse anti-LC3 (Cell Signaling Technology; Cat# 83506; 1:500), mouse anti-p62 (Biotechne; Cat# MAB8028; 2μg/mL), mouse anti-AMPKα (Cell Signaling Technology; Cat# 2793; 1:500), rabbit anti-phospho-AMPKα (T172) (Cell Signaling Technology; Cat# 2535; 1:500), mouse anti- GAPDH (Abcam; Cat# ab8245; 1:2000), rabbit anti-ATP6V1A (Cell Signaling Technology; Cat# 39517; 1:500), rabbit anti-NPC1 (Abcam; Cat# ab134113; 1:500), rabbit anti-VPS34 (Cell Signaling Technology; Cat# 4263; 1:500), rabbit anti-CatD (Cell Signaling Technology; Cat# 88239; 1:500), sheep anti-TREM2 (Biotechne; Cat# AF1729; 0.1μg/mL), rabbit anti-ATG7 (Cell Signaling Technology; Cat# 8558; 1:500), rabbit anti-ATG14 (Cell Signaling Technology; Cat# 96752; 1:500), and rabbit anti-UVRAG (Abcam; Cat# ab313627; 1:500).

Techniques: In Vitro, Expressing, Marker, Western Blot, Derivative Assay