Additional file 1 of Physical activity, energy expenditure and sedentary parameters in overfeeding studies - a systematic review

Risk of Bias Assessment. Risk of bias assessed for sequence generation, allocation concealment, blinding outcome assessors, incomplete outcome data, selective outcome and other sources of bias. (DOC× 133 kb

Expression of MEF2C, MYOD1 and MYOG inhibits SREBP-1 induced atrophy in myotubes.

<p>Human myotubes were infected for 48 h with recombinant adenoviruses encoding GFP, SREBP-1a or SREBP-1c, in combination with viruses expressing MEF2C, MYOD1, MYOG or GFP (control). (A) Representative images of myotube immunostaining with Myosin Heavy Chain antibody. (B) Myotube areas were measured as cell surface determination from MHC immunostaining pictures. The results are the means ± SE of 5 independent experiments (with 10 fields considered for each condition). *p<0.001 compared to Ad-GFP infected cells. #p<0.05 compared with Ad-SREBP-1a alone ; †p<0.05 compared with Ad-SREBP-1c alone; (C) Fusion index. The number of nuclei (total and in myotubes) was counted in all 12 conditions (with at least 4 fields for each condition). The results are the means ± SE of 3 independent experiments. *p<0.05 compared to Ad-GFP infected cells.</p

siRNA-mediated inhibition of SREBP-1 affects protein synthesis and degradation rates.

<p>Protein synthesis and degradation rates were determined in human primary myotubes under GFP or MRFs overexpression. Cells were transfected with siRNAs (control or against SREBP-1) and infected with recombinant adenovirus for 48 hours. The results are the means ± SE of 4 determinations. *p<0.05 compared to Ad-GFP infected cells. #p<0.05 compared to control siRNA.</p

Methodological approach.

<p>Methodological approach.</p

Expression of MEF2C, MYOD1 and MYOG are necessary to maintain muscle cell protein synthesis and expression of sarcomeric proteins.

<p>Human myotubes were infected for 48 h with recombinant adenoviruses encoding GFP (white), SREBP-1a (black) or SREBP-1c (grey) in combination with viruses expressing MEF2C, MYOD1, MYOG or GFP (control). (A) Protein synthesis rates were measured in all different conditions. The results are the means ± SE of 5 determinations. *p<0.001 compared to Ad-GFP infected cells. #p<0.05 compared with Ad-SREBP-1a alone ; †p<0.05 compared with Ad-SREBP-1c alone (B) Representative images of TNNI1 immuno-detection by western blotting from total cell extract of myotubes. (C) Quantification of TNNI1 content in each condition, normalized by tubulin and expressed as percentage of control. The results are the means ± SE of 3 determinations.</p

Opposite and indirect regulations of atrogenes by SREBP-1.

<p>Human myotubes were infected for 48 h with recombinant adenoviruses encoding GFP (white), SREBP-1a (black) or SREBP-1c (grey). (A) Protein levels of TRIM63 in myotubes overexpressing GFP, SREBP-1a or SREBP-1c. An illustrative immunoblot and quantification of the results are shown. The protein level of alpha-tubulin was used for normalization. The results presented are the means ± SE of 4 determinations. <b>*</b>p<0,05; <b>**</b>p<0.001 (B) TRIM63 proximal promoter activity on human myotubes. S represent Sterol Regulatory Element putative motif, E represent E-box motifs and M represent MRFs motifs. The results presented are the means of 5 determinations. (C) mRNA levels of atrogenic factors (FBXO32 and TRIM63) in response to GFP, SREBP-1a or SREBP-1c overexpression in combination with adenvirus-mediated overexpression of MEF2C, MYOD1, MYOG or GFP (control). The results presented are the means ± SE of 3 determinations. (D) Protein levels of tFOXO1 (total) and pFOXO1 (Ser256 phosphorylated) and tFOXO3 (total) and pFOXO3 (Ser253 phosphorylated) were determined by western blot experiments in myotubes overexpressing GFP, SREBP-1a or SREBP-1c. Immunodetection of alpha-tubulin was performed to normalize for the amount of loaded protein. Illustrative immunoblots are shown on the left, and the results of quantification are shown on the right. The results are presented as mean ± SE of 6 separate experiments. <b>*</b>p<0,05; <b>**</b>p<0.001.</p

Characteristics of the study population according to retirement status.

<p>Values are mean (SD) or %.</p><p>p values are from ANOVA or Chi-square tests.</p><p>Characteristics of the study population according to retirement status.</p

Sedentary behaviours and physical activity at baseline (2001) and changes during 6-year follow-up.

<p>Values are mean (SD) for baseline and mean (SE) for changes. p values are from ANOVA with adjustment on sex and age.</p>a<p>Comparisons between subjects not retired in 2001 and 2007 and subjects who retired between 2001 and 2007 are significantly different (p<0.05).</p>b<p>Comparisons between subjects who retired between 2001 and 2007 and subjects retired in 2001 and 2007 are significantly different (p<0.05).</p>c<p>Comparisons between subjects not retired in 2001 and 2007 and subjects retired in 2001 and 2007 are significantly different (p<0.05).</p>†<p>Except for vigorous leisure time physical activity and swimming, all values for changes are significantly different from 0.</p>††<p>All values for changes are significantly different from 0.</p>†††<p>Except for reading, domestic physical activity, vigorous leisure physical activity, walking, gardening and biking, all values for changes are significantly different from 0.</p><p>Sedentary behaviours and physical activity at baseline (2001) and changes during 6-year follow-up.</p

Scoping review’s flowchart.

<p>Scoping review’s flowchart.</p

Cross-tabulation of factors and currently used questionnaires.

<p>Cross-tabulation of factors and currently used questionnaires.</p