Effect of inhaled glucocortcoids and their metabolites on CYP3A5 mRNA expression in human lung cells

<p>File 1: Data are peak areas for the metabolites beclomethasone-17-monopropionate ([M1], the active drug) and Δ6-beclomethasone dipropionate ([M5], the CYP3A-mediated metabolite) and the internal standard (IS, prednisolone). Treatment groups were beclomethasone dipropionate (BDP) alone, beclomethasone dipropionate (BDP) plus esterase inhibitors (EI), 1-aminobenzotriazole (1-ABT) plus beclomethasone dipropionate (BDP), and 1-aminobenzotriazole (1-ABT) plus esterase inhibitors (EI) plus the active drug [M1]. Data corresponds to Figure 3A in the main text.</p> <p>File 2: Data are copy numbers of CYP3A5*1 mRNA and β-microglobulin (B2M, housekeeping gene) mRNA. Treatment groups are dimethyl sulfoxide (DMSO, vehicle control), beclomethasone dipropionate (BDP), beclomethasone dipropionate (BDP) plus esterase inhibitors (EI), 1-aminobenzotriazole (1-ABT), 1-aminobenzotriazole (1-ABT) plus beclomethasone dipropionate (BDP), and 1-aminobenzotriazole (1-ABT) plus esterase inhibitors (EI) plus active drug [M1]. Data corresponds to Figure 3B in the main text. </p> <p>File 3: Data are copy numbers of CYP3A5*1 mRNA and β-microglobulin (B2M, housekeeping gene) mRNA. Treatment groups were dimethyl sulfoxide (DMSO, vehicle control), beclomethasone dipropionate (BDP), ketoconazole (KTZ) at 50 µM plus beclomethasone dipropionate (BDP), ketoconazole (KTZ) at 10 µM plus beclomethasone dipropionate (BDP), and ketoconazole (KTZ) at 1 µM plus beclomethasone dipropionate (BDP). Data corresponds to Figure 4A in the main text. </p> <p>File 4: Data are copy numbers of CYP3A5*1 mRNA and β-microglobulin (B2M, housekeeping gene) mRNA. Treatment groups were dimethyl sulfoxide (DMSO, vehicle control), budesonide (BUD), ketoconazole (KTZ) at 50 µM plus budesonide (BUD), ketoconazole (KTZ) at 10 µM plus budesonide (BUD), and ketoconazole (KTZ) at 1 µM plus budesonide (BUD). Data corresponds to Figure 4B in the main text.</p> <p>File 5: Data are copy numbers of CYP3A5*1 mRNA and β-microglobulin (B2M, housekeeping gene) mRNA. Treatment groups were dimethyl sulfoxide (DMSO, vehicle control), triamcinolone acetonide (TCL), ketoconazole (KTZ) at 50 µM plus triamcinolone acetonide (TCL), ketoconazole (KTZ) at 10 µM plus triamcinolone acetonide (TCL), and ketoconazole (KTZ) at 1 µM plus triamcinolone acetonide (TCL). Data corresponds to Figure 4C in the main text. </p> <p>File 6: Data are copy numbers of CYP3A5*1 mRNA and β-microglobulin (B2M, housekeeping gene) mRNA. Treatment groups were dimethyl sulfoxide (DMSO, vehicle control), fluticasone propionate (FLT), ketoconazole (KTZ) at 50 µM plus fluticasone propionate (FLT), ketoconazole (KTZ) at 10 µM plus fluticasone propionate (FLT), and ketoconazole (KTZ) at 1 µM plus fluticasone propionate (FLT). Data corresponds to Figure 4D in the main text.</p> <p>File 7: Data are copy numbers of CYP3A5*1 mRNA and β-microglobulin (B2M, housekeeping gene) mRNA. Treatment groups were dimethyl sulfoxide (DMSO, vehicle control), flunisolide (FLN), ketoconazole (KTZ) at 50 µM plus flunisolide (FLN), ketoconazole (KTZ) at 10 µM plus flunisolide (FLN), and ketoconazole (KTZ) at 1 µM plus flunisolide (FLN). Data corresponds to Figure 4E in the main text. </p> <p>File 8: Data are copy numbers of CYP3A5*1 mRNA and β-microglobulin (B2M, housekeeping gene) mRNA. Control treatment groups were ketoconazole (KTZ) at 50 µM, ketoconazole (KTZ) at 10 µM, and ketoconazole (KTZ) at 1 µM in the main text.</p> <p>File 9: Data are represented as copies of mRNA detected for the glucocorticoid receptor (GR) and β-microglobulin (B2M, housekeeping gene) for each treatment group. siRNA directed against the green fluorescent protein (GFP) was used as a negative control and compared to siRNA directed against the glucocorticoid receptor (GR) at 48, 72, and 96 hr post siRNA treatment. Data corresponds to Figure 5A in the main text. </p> <p>File 10: Data are copy numbers of mRNA detected for CYP3A5*1 and β-microglobulin (B2M, housekeeping gene). Pretreatments were siRNA directed against either green fluorescent protein (GFP, negative control) or the glucocorticoid receptor (GR). Three treatment groups consisted of dimethyl sulfoxide (DMSO, vehicle control), beclomethasone dipropionate (BDP), and beclomethasone dipropionate (BDP) plus esterase inhibitors (EI). Data corresponds to Figure 5B in the main text. </p> <p>File 11: Data are copies of CY3A5*1 mRNA divided by 10,000 copies of β-microglobulin (B2M, housekeeping gene) for each treatment. Treatments were dimethyl sulfoxide (DMSO, vehicle control), beclomethasone dipropionate (BDP), dimethyl sulfoxide (DMSO, vehicle control) treated cells cultured without hydrocortisone (HC), and beclomethasone dipropionate (BDP) treated cells cultured without hydrocortisone (HC). All treatments lasted for 24 hours. Data corresponds to Figure 6 in the main text. </p>