Age- and BMI-Associated Expression of Angiogenic Factors in White Adipose Tissue of Children

Figures

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  Figure A1. Association of adipocyte diameter and VWF expression with age and BMI-SDS in the WAT of infants <6 and children ≥6 months. Adipocyte size was determined in histologic sections as specified in Methods. Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using a specific primer pair as indicated. VWF gene expression was calculated as described in Methods and plotted against age. (A) In contrast to the analysis with all children, adipocyte size did not correlate positively with age in the infants <6 months. (C) Adipocyte size did, in turn, correlated positively with BMI-SDS. (B,D) VWF correlated positively with age and BMI-SDS in the infants <6 months. (E–F) There were no significant correlations of adipocyte diameter and VWF with age or BMI-SDS in the children ≥6 months. Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.

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  Figure A2. Association of angiogenic gene expression with age in the WAT of infants <6 and children ≥6 months. Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using specific primer pairs as indicated. Respective gene expression was calculated as described in Methods and plotted against age. (A) As in the correlation analysis with all children, VEGFA expression showed no significant correlation with age in the infants <6 months. (B,C) Both FGF1 and FGF2 expression still correlated positively with age when performing the analyses with the infants younger than 6 months only. (D) ANGPT1 showed no significant correlation with age. (E,F) In contrast to the analysis with all children, ANGPT2 showed no significant correlation with age (E), while TEK correlated positively with age in the infants <6 months. (F). (G–L) There were no significant correlations of the analyzed angiogenic factors with age in the group of children ≥6 months. Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.

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  Figure A3. Association of angiogenic gene expression with BMI-SDS in the WAT of infants <6 and children ≥6 months. Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using specific primer pairs as indicated. Respective gene expression was calculated as described in Methods and plotted against BMI-SDS. (A–F) Like in the analysis with all children, ANGPT1 and the receptor TEK correlated positively with BMI-SDS in the infants <6 months (D,F), while VEGFA, FGF1, FGF2, and ANGPT2 showed no significant correlation (A–C,E). (G–L) We found no significant correlation of angiogenic factors with BMI-SDS in the children ≥6 months. Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.

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  Figure A4. Association of angiogenic gene expression with adipocyte size in the WAT of infants <6 and children ≥6 months. Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using specific primer pairs as indicated. Respective gene expression was calculated as described in Methods and plotted against adipocyte diameter. (B) Only FGF1 correlated positively with adipocyte size in the infants <6 months. (C,D) Both FGF2 and ANGPT1, which correlated positively with adipocyte diameter in the analysis with all children, did not correlate significantly with adipocyte size in the infants <6 months. (A,E,F) VEGFA, ANGPT2, and TEK showed no significant correlations with adipocyte diameter in the <6 months group. (G–L) In contrast, in the children ≥6 months, only ANGPT1 correlated positively with adipocyte diameter (J), while there was no correlation of the other angiogenic factors (G–I,K,L). Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.

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  Figure A5. Association of VWF with angiogenic gene expression in the WAT of infants <6 and children ≥6 months. Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using specific primer pairs as indicated. Respective gene expression was calculated as described in Methods. (A–F) Like in the analysis with all children, all analyzed angiogenic factors, with the exception of ANGT2, correlated positively with VWF when performing the analysis with the infants <6 months only. (G–L) In the children ≥6 months VEGFA did not correlate significantly with VWF (G). In contrast to the analyses with the infants <6 months and all children, ANGPT2 correlated positively with VWF in the ≥6 months group (K). (H–J,L) As in the infants <6 months FGF1, FGF2, ANGPT1, and L also correlated positively with VWF in the children ≥6 months. Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.

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  Figure 1. Association of adipocyte size and PDGFRa expression with age and BMI-SDS. (A,B) Pictures of representative adipose tissue sections stained with haematoxylin and eosin from a patient with small (A) and a patient with large (B) adipocytes. (C,D) Adipocyte diameter was plotted against age (C) and BMI-SDS (D). Adipocyte size correlated positively with age and BMI-SDS in our collective (C,D). (E,F) Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using specific primer pairs as indicated. PDGFRa gene expression was calculated as described in Methods and plotted against age. PDGFRa expression correlated positively with age (E) but not with BMI-SDS (F) in the collective. Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.

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  Figure 2. Association of angiogenic gene expression in pediatric WAT with age. Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using specific primer pairs as indicated. Respective gene expression was calculated as described in Methods and plotted against age. (A) VEGFA expression showed no significant correlation with age. (B,C) FGF1 and FGF2 expression, however, correlated positively with age. (D–F) While ANGPT1 and the receptor TEK showed no significant correlation with age (D,F), ANGPT2 displayed a negative correlation (E). Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.

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  Figure 3. Association of angiogenic gene expression in pediatric WAT with BMI-SDS. Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using specific primer pairs as indicated. Respective gene expression was calculated as described in Methods and plotted against BMI-SDS. (D,F) ANGPT1 and the receptor TEK correlated positively with BMI-SDS. (A–C,E) In contrast, VEGFA, FGF1, FGF2, and ANGPT2 showed no significant correlation. Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.

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  Figure 4. Association of angiogenic gene expression in pediatric WAT with adipocyte size. Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using specific primer pairs as indicated. Respective gene expression was calculated as described in Methods and plotted against adipocyte diameter. (B–D) FGF1, FGF2, and ANGPT1 showed a positive correlation with adipocyte diameter. (A,E,F) VEGFA, ANGPT2, and TEK, in contrast, showed no significant correlations with adipocyte diameter. Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.

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  Figure 5. Association of angiogenic gene expression in pediatric WAT with VWF expression. Total RNA was isolated, reverse transcribed, and subjected to qPCR analysis using specific primer pairs as indicated. Respective gene expression was calculated as described in Methods. (A–D) VEGFA, FGF1, FGF2, and ANGPT1 correlated positively with VWF. (E) There was no significant correlation between ANGPT2 and VWF. (F) TEK, in turn, showed the strongest correlation with VWF. Spearman correlation coefficient rs and p value, as well as number of subjects, are given in each scatter plot. Significant p values (p < 0.05) are indicated in bold. Trend lines were calculated as linear regressions.