anti cd36  (R&D Systems)

Journal: Channels

Article Title: Inhibition of lipolysis in visceral adipose tissue from obese mice and humans prevents impairment of endothelial Kir2.1 channels

doi: 10.1080/19336950.2025.2564651

Figure Lengend Snippet: Inhibition of lipolysis in VAT of obese mice prevents impairment of endothelial Kir2.1. Human adipose microvascular endothelial cells (HAMECs) were incubated with either 200 μM 6:1 palmitic acid (PA):BSA or adipose tissue from lean or obese mice (en bloc; 5 mg/ml) for 48 h before assessing inwardly rectifying K + currents via whole cell patch clamp. The holding potential was −30 mV, and the voltage ramp protocol was from − 140 to +40 mV over 400 ms. The representative recordings shown in A, C, and E reveal that the inwardly rectifying K + currents reverse near the predicted reversal potential of approximately −21 mV under the specified patch conditions (i.e. 60 mM K + bath, 140 mM K + pipette solutions). A) Representative current recordings from untreated control HAMECs and HAMECs treated with either vehicle control (FA-free BSA) or PA. B) Group data show normalized current densities (pA/pF) analyzed and compared at −100 mV to determine the effects of PA treatment on Kir2.1 function ( n = 15–17 cells/group tested over three independent experiments). Significant differences (*) were determined by a Kruskal-Wallis test ( p = 0.0328) followed by Dunn’s post hoc tests. There was no difference in cell capacitance among groups (control, 46.5 ± 3.4 pF; vehicle, 46.0 ± 2.7 pF; FA, 52.2 pF ±4.4 pF; p = 0.5487). C) Representative current recordings from HAMECs treated with PA following pretreatment of cells with scrambled control (scr con) or CD36 siRNA. D) Group data show normalized current densities (pA/pF) analyzed and compared at −100 mV to determine the effects of CD36 downregulation on Kir2.1 function in the presence of PA ( n = 14–16 cells/group tested over 3 independent experiments). A significant difference (*) relative to the scrambled control (scr con) was determined using a Mann-Whitney test. There was no difference in cell capacitance between groups (scrambled control, 64.8 ± 4.8 pF; CD36 siRNA, 62.3 ± 4.8 pF; p = 0.8773). E) Representative current recordings from untreated control cells (no AT) or cells exposed to VAT from obese mice pretreated with either DMSO vehicle (vehicle) or 20 μM Atglistatin. F) Group data reveal normalized current densities (pA/pF) that were analyzed and compared at −100 mV to determine the effects of treating VAT with Atglistatin on endothelial Kir2.1 function ( n = 20–25 cells/group). VAT from four mice (two males, two females) was used across four independent experiments. Significant differences (*) were determined by a Kruskal-Wallis test ( p = 0.0004) followed by Dunn’s post hoc tests. There was no difference in cell capacitance among groups (control, 56.6 pF ±4.8; vehicle, 51.8 ± 4.5 pF; Atglistatin, 60.7 ± 4.0 pF; p = 0.2649).

Article Snippet: Following blocking, cells were washed twice with PBS for 10 min. Primary antibodies targeting extracellular epitopes of CD36 (R&D Systems, polyclonal goat anti-mouse CD36, Cat. No. AF2519) or Kir2.1 (Abcam, rabbit monoclonal; Cat. No. AB109750 ) was diluted 1:100 in PBS containing 1% BSA.

Techniques: Inhibition, Incubation, Patch Clamp, Transferring, Control, MANN-WHITNEY

Journal: Channels

Article Title: Inhibition of lipolysis in visceral adipose tissue from obese mice and humans prevents impairment of endothelial Kir2.1 channels

doi: 10.1080/19336950.2025.2564651

Figure Lengend Snippet: VAT does not affect endothelial Kir2.1 or CD36 expression. HAMECs were incubated en bloc (5 mg/ml) with adipose tissue from lean or obese mice or human subjects with obesity for 48 hours before assessing expression of Kir2.1 or CD36 using immunocytochemistry. HAMECs that were not exposed to adipose served as a control (no AT). A) representative fluorescent microscopy images (10x) of HAMECs showing expression of Kir2.1 after exposure to VAT from lean or obese mice (top). Nuclei were stained with DAPI (bottom). The scale bar is 20 μm. B) normalized group data reveal no differences in mean fluorescence intensity (FI) as determined by a Kruskal-wallis test ( p = 0.1105) ( n = 7). C) representative fluorescent microscopy images (10x) of HAMECs showing expression of CD36 after exposure to VAT from lean or obese mice (top). Nuclei were stained with DAPI (bottom). The scale bar is 20 μm. D) normalized group data reveal no differences in mean FI as determined by a Kruskal-wallis test ( p = 0.4834) ( n = 7). E) representative fluorescent microscopy images (10x) of HAMECs showing expression of Kir2.1 after exposure to SAT or VAT from human subjects (top). Nuclei were stained with DAPI (bottom). The scale bar is 20 μm. F) normalized group data reveal no differences in mean FI as determined by a Kruskal-wallis test ( p = 0.0866) ( n = 4). G) representative fluorescent microscopy images (10x) of HAMECs showing expression of CD36 after exposure to SAT or VAT from human subjects (top). Nuclei were stained with DAPI (bottom). The scale bar is 20 μm. H) normalized group data reveal no differences in mean FI as determined by a Kruskal-wallis test ( p = 0.4121) ( n = 4).

Article Snippet: Following blocking, cells were washed twice with PBS for 10 min. Primary antibodies targeting extracellular epitopes of CD36 (R&D Systems, polyclonal goat anti-mouse CD36, Cat. No. AF2519) or Kir2.1 (Abcam, rabbit monoclonal; Cat. No. AB109750 ) was diluted 1:100 in PBS containing 1% BSA.

Techniques: Expressing, Incubation, Immunocytochemistry, Control, Microscopy, Staining, Fluorescence