Figures
- Fig. 1—
Cross-sectional area (CSA) of type I and type II fibres in the diaphragm of six controls (▓;) and six patients with severe chronic obstructive pulmonary disease (□). Data are presented as mean±sd. *: p<0.05.
- Fig. 2—
a) MyoD mRNA (expressed per 106 18s copies) and b, c) protein (relative to controls) expression in the diaphragm of controls and patients with chronic obstructive pulmonary disease (COPD). For mRNA, each data point represents an individual patient and horizontal lines represent the mean value. a) Controls: n = 10; COPD: n = 11. b, c) Controls: n = 9; COPD: n = 15. *: p<0.05; #: p<0.005.
- Fig. 3—
a) MAFbx mRNA expression and b) Nedd4 mRNA expression in the diaphragm of controls (n = 10) and patients with chronic obstructive pulmonary disease (COPD; n = 11). mRNA is expressed per 106 18s copies. Each data point represents an individual patient and the horizontal lines represent the mean value. *: p<0.05. #: p<0.005.
- Fig. 4—
a) Nuclear factor (NF)-κB p50 DNA-binding activity and protein expression of inhibitor proteins b) IκBα (35 kDa) and c) IκBβ (45 kDa) (normalised to α-tubulin (55 kDa) and expressed relative to controls) in the diaphragm of controls (n = 9) and patients with chronic obstructive pulmonary disease (COPD; n = 15). Data are presented as mean±sd. **: p<0.01.
- Fig. 5—
a) Myostatin mRNA (expressed per 106 18S copies) and myostatin protein levels (26 kDa) in b) cytoplasmic and c) nuclear fraction in the diaphragm of controls and patients with chronic obstructive pulmonary disease (COPD). Each data point in a) represents an individual patient. Controls: n = 10; COPD: n = 11. Horizontal lines represent the mean value. a, c) Controls: n = 9; COPD: n = 15. Data normalised to α-tubulin (c; 55 kDa) expressed relative to controls presented as mean±sd. Con: control. *: p<0.05.
Tables
- Table 1—
Anthropometric characteristics and pulmonary function tests in the controls and in the patients with chronic obstructive pulmonary disease(COPD)
Characteristics Controls COPD Subjects n 13 19 Male/female n 11/2 8/11 Age yrs 64±3 58±1 Weight kg 71±3 58±2** BMI kg·m-2 25±1 21±1** FEV1 L 2.67±0.21 0.71±0.05# FEV1 % pred 98±5 26±2# FEV1/FVC % 76±2 33±2# TLC % pred 95±6 126±5*** RV % pred 107±9 227±12¶ DL,CO % pred 83±5 29±2# KCO % pred 105±6 44±3# Data are presented as mean±sem, unless otherwise stated. BMI: body mass index; FEV1: forced expiratory volume in 1 s; % pred: % predicted; FVC: forced vital capacity; TLC: total lung capacity; RV: residual volume; DL,CO: diffusing capacity of the lung for carbon monoxide; KCO: DL,CO per unit of alveolar volume. #: p<0.0001; ¶: p<0.00005; **: p<0.01; ***: p<0.001.
- Table 2—
Primer sequences used for real-time PCR
MAFbx Forward: 5′ GTG GTA CTG AAA GTC CTT GAA GAC 3′Reverse: 5′ TTA ATG TTC CCG ACC AGC A 3′ MuRF1 Forward: 5′ GAA TAA CTG TAT CTC CAT GCT GG 3′Reverse: 5′ GGC ATA CAA CGT GTC AAA CTT 3′ MyoD Forward: 5′ GAC GGC ATG ATG GAC TAC AG 3′Reverse: 5′ AGG CAG TCT AGG CTC GAC AC 3′ Myogenin Forward: 5′ AGC GAA TGC AGC TCT CAC AG 3′Reverse: 5′ AGG TTG TGG GCA TCT GTA GG 3′ Nedd4 Forward: 5′ TTT GGA AAT TCA GCC GTG AG 3′Reverse: 5′ CCT GGT GGT AAT CCA GAT GAA 3′ Myostatin Forward: 5′ TGT AAC CTT CCC AGG ACC AG 3′Reverse: 5′ GGT AAC GAC AGC ATC GTG ATT 3′
Supplementary methods and figures
Files in this Data Supplement:
- Supplementary methods and figures - Methods: histochemistry; RNA extraction and real-time quantitative PCR; protein extraction; gel electrophoresis and immunoblotting; NF-κB activity; Figure E1. Representative blots stained with PonceauS, illustrating equal loading for MyoD protein (A), myogenin protein (B) and Myostatin nuclear protein (C) in the diaphragm of 6 different controls and 6 different COPD patients. Figure E2. Western blot illustrating Histone-H3 (16 kDa) expression in the diaphragm of 6 different controls and 6 different COPD patients.
