Characterization of TGF-β expression and signaling profile in the adipose tissue of rats fed with high-fat and energy-restricted diets Bernardo Sousa-Pinto,

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Characterization of TGF-β expression and signaling profile in the adipose tissue of rats fed with high-fat and energy-restricted diets Bernardo Sousa-Pinto, Laura Gonçalves, Adriana R. Rodrigues, Inês Tomada, Henrique Almeida, Delminda Neves, Alexandra M. Gouveia Journal of Nutritional Biochemistry Volume 38, Pages 107-115 (December 2016) DOI: 10.1016/j.jnutbio.2016.07.017 Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 1 mRNA levels of TGF-β/actin (A), TGF-βRI/actin (B) and TGF-βRII/actin (C) in the mesenteric, epididymal, retroperitoneal and subcutaneous AT from control, HFD or ER rats. Asterisks (*) indicate groups that are statistically different from controls using two-tailed Student's t test (*P<.05). Hashes (#) indicate control fat depots that are statistically different from mesenteric fat using ANOVA followed by Tukey's correction test. Dollar signs ($) indicate fat depots that are statistically different from subcutaneous fat of rats of the same diet group using ANOVA followed by Tukey's correction test. Quotation marks (") indicate control diet fat depots that are statistically different from retroperitoneal fat using ANOVA followed by Tukey's correction test. Journal of Nutritional Biochemistry 2016 38, 107-115DOI: (10.1016/j.jnutbio.2016.07.017) Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 2 Western blot analysis of TGF-β, IL-1β, IL-10, Smad3, phospho-Smad3, Akt, phospho-Akt, ERK1/2, phospho-ERK1/2 and MAPKp38, in mesenteric AT from all experimental groups, as well as representative Ponceau stain (A). The graphs represent the semiquantitative analysis, calculated by the fraction of pixels presented by each band relative to the Ponceau S staining (B). Error bars represent standard error of the mean. Asterisks indicate groups that are statistically different from controls using two-tailed Student's t test (*P<.05). Journal of Nutritional Biochemistry 2016 38, 107-115DOI: (10.1016/j.jnutbio.2016.07.017) Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 3 Western blot analysis of TGF-β, IL-1β, IL-10, Smad3, phospho-Smad3, Akt, phospho-Akt, ERK1/2, phospho-ERK1/2 and MAPKp38, in epididymal AT from all experimental groups, as well as representative Ponceau stain (A). The graphs represent the semiquantitative analysis, calculated by the fraction of pixels presented by each band relative to the Ponceau S staining (B). Error bars represent standard error of the mean. Asterisks indicate groups that are statistically different from controls using two-tailed Student's t test (*P<.05). Journal of Nutritional Biochemistry 2016 38, 107-115DOI: (10.1016/j.jnutbio.2016.07.017) Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 4 Western blot analysis of TGF-β, IL-1β, IL-10, Smad3, phospho-Smad3, Akt, phospho-Akt, ERK1/2, phospho-ERK1/2 and MAPKp38, in retroperitoneal AT from all experimental groups, as well as representative Ponceau stain (A). The graphs represent the semiquantitative analysis, calculated by the fraction of pixels presented by each band relative to the Ponceau S staining (B). Error bars represent standard error of the mean. Asterisks indicate groups that are statistically different from controls using two-tailed Student's t test (*P<.05). Journal of Nutritional Biochemistry 2016 38, 107-115DOI: (10.1016/j.jnutbio.2016.07.017) Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 5 Western blot analysis of TGF-β, IL-1β, IL-10, ERK1/2 and MAPK p38, in subcutaneous AT from all experimental groups, as well as representative Ponceau stain (A). The graphs represent the semiquantitative analysis, calculated by the fraction of pixels presented by each band relative to the Ponceau S staining (B). Error bars represent standard error of the mean. Asterisks indicate groups that are statistically different from controls using two-tailed Student's t test (*P<.05). Journal of Nutritional Biochemistry 2016 38, 107-115DOI: (10.1016/j.jnutbio.2016.07.017) Copyright © 2016 Elsevier Inc. Terms and Conditions

Fig. 6 mRNA levels of PAI-1/actin (A) and GLUT4/actin (B) mRNA levels in the mesenteric, epididymal, retroperitoneal and subcutaneous AT from control, HFD or ER rats. Asterisks (*) indicate groups that are statistically different from controls using two-tailed Student's t test (*P<.05). Hashes (#) indicate control fat depots that are statistically different from mesenteric fat using ANOVA followed by Tukey's correction test. Quotation marks (") indicate HFD fat depots that are statistically different from retroperitoneal fat using ANOVA followed by Tukey's correction test. Journal of Nutritional Biochemistry 2016 38, 107-115DOI: (10.1016/j.jnutbio.2016.07.017) Copyright © 2016 Elsevier Inc. Terms and Conditions

Journal of Nutritional Biochemistry 2016 38, 107-115DOI: (10. 1016/j Journal of Nutritional Biochemistry 2016 38, 107-115DOI: (10.1016/j.jnutbio.2016.07.017) Copyright © 2016 Elsevier Inc. Terms and Conditions