Figure S4 E. (F) Immunoblots show pHSL, HSL, and ATGL expression in eWAT at 7DPI. GAPDH is used as a loading control. (G) Quantification of (F). Mean and SEM (unpaired two-tailed Student’s t test ∗∗ p < 0.01, AAV-GFP/SCI n = 4 and AAV-Cre/SCI n = 4 biological replicates). (H) CGRP concentration in eWAT at 7DPI. Mean and SEM (unpaired two-tailed Student’s t test ∗∗ p < 0.01, AAV-GFP/SCI n = 4 and AAV-Cre/SCI n = 4 biological replicates). (I) Representative hematoxylin and eosin images of eWAT at 7DPI. Scale, 100 μm. (J) Quantification of adipocyte area in (I). Mean and SEM (two-way ANOVA ∗∗∗ p < 0.001, ns [not significant], Na v 1.8-Cre n = 5 and Na v 1.8-Cre/ Cacna2d1 fl/fl n = 5 biological replicates; 2,732–3,170 adipocytes/experimental condition). (K) Ex vivo eWAT lipolysis at 7DPI. Mean and SEM (unpaired two-tailed Student’s t test ∗ p < 0.05, Na v 1.8-Cre n = 4 and Na v 1.8-Cre/ Cacna2d1 fl/fl n = 5 biological replicates). (L) Immunoblots show pHSL, HSL, and ATGL expression in eWAT at 7DPI. GAPDH is used as a loading control. (M) Quantification of (L). Mean and SEM (unpaired two-tailed Student’s t test ∗ p < 0.05, Na v 1.8-Cre n = 4 and Na v 1.8-Cre/ Cacna2d1 fl/fl n = 5 biological replicates). (N) CGRP concentration in eWAT at 7DPI. Mean and SEM (unpaired two-tailed Student’s t test ∗∗ p < 0.01, Na v 1.8-Cre n = 8 and Na v 1.8-Cre/ Cacna2d1 fl/fl n = 10 biological replicates). " width="100%" height="100%">
Journal: Cell Reports Medicine
Article Title: α2δ1-mediated maladaptive sensory plasticity disrupts adipose tissue homeostasis following spinal cord injury
doi: 10.1016/j.xcrm.2024.101525
Figure Lengend Snippet: Cacna2d1 conditional deletion in DRG neurons normalizes eWAT lipolysis after SCI (A) Experimental scheme and timeline of AAV injection to transduce L3–5 DRG. (B) Real-time qPCR shows Cacna2d1 normalized expression in adult L3–5 DRG from Cacna2d1 fl/fl mice injected with either AAV-GFP or AAV-Cre. Mean and SEM (unpaired two-tailed Student’s t test ∗ p < 0.05, AAV-Ctr n = 4 and AAV-Cre n = 4 biological replicates). (C) Representative hematoxylin and eosin images of eWAT at 7 days post injury (DPI). Scale, 100 μm. (D) Quantification of adipocyte area in (C). Mean and SEM (two-way ANOVA ∗∗ p < 0.01 and ∗∗∗ p < 0.001, ns [not significant], AAV-GFP/SCI n = 3 and AAV-Cre/SCI n = 3 biological replicates; 2,063–2,322 adipocytes/experimental condition). (E) Ex vivo eWAT lipolysis at 7DPI. Mean and SEM (Kruskal-Wallis test followed by Dunn’s multiple comparisons test ∗∗ p < 0.01, ns [not significant], AAV-GFP n = 6, AAV-GFP/SCI n = 5, and AAV-Cre/SCI n = 6 biological replicates). The AAV-GFP samples are the same as in Figure S4 E. (F) Immunoblots show pHSL, HSL, and ATGL expression in eWAT at 7DPI. GAPDH is used as a loading control. (G) Quantification of (F). Mean and SEM (unpaired two-tailed Student’s t test ∗∗ p < 0.01, AAV-GFP/SCI n = 4 and AAV-Cre/SCI n = 4 biological replicates). (H) CGRP concentration in eWAT at 7DPI. Mean and SEM (unpaired two-tailed Student’s t test ∗∗ p < 0.01, AAV-GFP/SCI n = 4 and AAV-Cre/SCI n = 4 biological replicates). (I) Representative hematoxylin and eosin images of eWAT at 7DPI. Scale, 100 μm. (J) Quantification of adipocyte area in (I). Mean and SEM (two-way ANOVA ∗∗∗ p < 0.001, ns [not significant], Na v 1.8-Cre n = 5 and Na v 1.8-Cre/ Cacna2d1 fl/fl n = 5 biological replicates; 2,732–3,170 adipocytes/experimental condition). (K) Ex vivo eWAT lipolysis at 7DPI. Mean and SEM (unpaired two-tailed Student’s t test ∗ p < 0.05, Na v 1.8-Cre n = 4 and Na v 1.8-Cre/ Cacna2d1 fl/fl n = 5 biological replicates). (L) Immunoblots show pHSL, HSL, and ATGL expression in eWAT at 7DPI. GAPDH is used as a loading control. (M) Quantification of (L). Mean and SEM (unpaired two-tailed Student’s t test ∗ p < 0.05, Na v 1.8-Cre n = 4 and Na v 1.8-Cre/ Cacna2d1 fl/fl n = 5 biological replicates). (N) CGRP concentration in eWAT at 7DPI. Mean and SEM (unpaired two-tailed Student’s t test ∗∗ p < 0.01, Na v 1.8-Cre n = 8 and Na v 1.8-Cre/ Cacna2d1 fl/fl n = 10 biological replicates).
Article Snippet: Mouse CGRP ELISA kit (colorimetric) , Novus , Cat #NBP3-00522.
Techniques: Injection, Transduction, Expressing, Two Tailed Test, Ex Vivo, Western Blot, Control, Concentration Assay