Additional file 1 of Endothelial cell regulation of systemic haemodynamics and metabolism acts through the HIF transcription factors
Creators: name: Lambden, Simon
name: Cowburn, Andrew S.
name: Macias, David
name: Garrud, Tessa A. C.
name: Krause, Bernardo J.
name: Giussani, Dino A.
name: Summers, Charlotte
name: Johnson, Randall S.
Publisher: name: figshare
Year: 2021
Subjects: subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Physiology
subject: Science Policy
subject: Immunology
subject: FOS: Clinical medicine
subject: 69999 Biological Sciences not elsewhere classified
subject: Marine Biology
subject: 111714 Mental Health
subject: FOS: Health sciences
subject: Computational Biology

description: Additional file 1: Figure S1. Impact of global and pulmonary endothelial specific HIF-1α knockout on the right heart and pulmonary vasculature. A: Right ventricular systolic pressure (RVSP) was measured under isoflurane anaesthesia in 21% oxygen in HIF-1α Tie2 Cre (green, n = 3) and HIF-1α flox/flox littermates (grey, n = 3). Data presented as mean ± SD, analysis by unpaired t test p = 0.77. B: Fulton index of hearts dissected from HIF-1α Tie2 Cre (green, n = 4) and HIF-1α flox/flox littermates (grey, n = 4). Data presented as mean ± SD, analysis by unpaired t test p = 0.78. C: Pulmonary vascular remodelling was determined in HIF-1α Tie2 Cre (green, n = 4) and HIF-1α flox/flox littermates (grey, n = 4). Quantification of the parabronchial medial thickness on smooth muscle actin stained vessels is presented as a percentage of vessel wall thickness, analysis by unpaired t test, p = 0.33. D: Representative images of parabronchial vessels from HIF-1α Tie2 Cre mice stained with (i) haematoxylin and eosin (H&amp;E), (ii) smooth muscle actin (SMA), (iii) Elastic tissue fibres—Verhoeff’s Van Gieson (EVG). (iv) Representative image of microvasculature of HIF-1α Tie2 Cre stained for SMA. E: Right ventricular systolic pressure (RVSP) was measured under isoflurane anaesthesia in 21% oxygen in HIF-1α L1 Cre (Red, n = 5) and HIF-1α flox/flox littermates (grey, n = 7). Data presented as mean ± SD, analysis by two-way ANOVA p = n0.99. F: Fulton index of hearts dissected from HIF-1α L1 Cre (Red, n = 6) and HIF-1α flox/flox littermates (grey, n = 7). Data presented as mean ± SD, analysis by unpaired t Test p = 0.66. G: pulmonary vascular remodelling was determined in HIF-1α L1Cre (red, n = 5) and HIF-1α flox/flox littermates (grey, n = 5). Quantification of the parabronchial intimal medial thickness on smooth muscle actin stained vessels is presented as a percentage of vessel wall thickness, analysis by unpaired t test, p = 0.62. H: Representative images of parabronchial vessels from HIF-1α L1 Cre mice stained with (i) H&amp;E, (ii) SMA, (iii) EVG. (iv) Representative image of microvasculature of HIF-1α Tie2 Cre stained for SMA. Figure S2. Haematological, Biochemical and Cytokine analysis of pulmonary endothelial HIF-1α knockout mice (HIF-1α L1Cre, red) and HIF-1α flox/flox littermates (grey). A: Haemoglobin concentration (g/dL) and, B: Red Blood Cell count (RBC × 106 /mm3) in knockout and wild-type littermate controls (n = 7, p = ns). C: Renal function measured by plasma creatinine and urea in knockout (n = 5) and wild-type littermate controls (n = 6, p = ns). D: Salt handling measured by analysis of plasma sodium and chloride concentrations in knockout (n = 5) and wild-type littermate controls (n = 6, p = ns). E and F: Circulating plasma cytokine concentrations measured using a multiplex panel in knockout and wild-type littermates, n = 5, all p = ns. Figure S3. Effects of HIF-2α pulmonary endothelial knockout on constitutive cardiovascular function. Circadian variations in A: Systolic, B: Diastolic blood pressure, C: heart rate, D: subcutaneous temperature, E: VO2 and F: VCO2 of HIF-2α L1 Cre (Blue, n = 4) and littermate HIF-1α flox/flox (grey, n = 4) mice were recorded by radio-telemetry. Black box represents nocturnal phase. Data are presented as a mean ± SEM for each 30 min period, p values for area under the curve followed by unpaired t test are shown. G: Quantification of the parabronchial medial thickness on smooth muscle actin stained vessels is presented as a percentage of vessel wall thickness, analysis by unpaired t test, p = ns. Figure S4. Effects of HIF-2α pulmonary endothelial knockout on response to acute hypoxia. Impact of acute hypoxia with inspired oxygen concentration of 11% on A: systolic (p = 0.91), B: diastolic blood pressure (p