Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: Primers for real-time PCR.

Article Snippet: The miR-548au-3p mimics, inhibitor and negative control (NC), CA12 overexpression plasmid, and small interfering RNA (siRNA) targeting CA12 and siRNA NC were provided by Guangzhou RiboBio Co., Ltd. and transfected into rat tracheal chondrocytes.

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: Identification of DEGs involved in the pathogenesis of CPAM. (a) Flowchart of the study design and samples at each stage of analysis. (b) Scatterplot of mRNA expression variation between diseased CPAM and normal tissues. (c) Hierarchical cluster of gene expression profiles from microarray assays. Expression of CA12, LONRF3, MAP2, THBS1, and PPID at mRNA (d) and protein (e) levels in lung tissues from CPAM and adjacent normal tissues. ∗∗∗ p < 0.001, compared with normal.

Article Snippet: The miR-548au-3p mimics, inhibitor and negative control (NC), CA12 overexpression plasmid, and small interfering RNA (siRNA) targeting CA12 and siRNA NC were provided by Guangzhou RiboBio Co., Ltd. and transfected into rat tracheal chondrocytes.

Techniques: Expressing, Gene Expression, Microarray

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: CA12 as a direct target of miR-548au-3p. (a) Binding sites between miR-548au-3p and the 3′-UTR region of CA12 mRNA. (b) HEK-293 cells were cotransfected with miR-548au-3p mimics or NC and wild-type or mutant CA12 3′-UTR, followed by luciferase reporter assay. CA12 mRNA (c) and protein (d) levels in rat tracheal chondrocytes after transfection with miR-548au-3p mimics or inhibitor. ∗∗ p < 0.01 and ∗∗∗ p < 0.001, compared with NC.

Article Snippet: The miR-548au-3p mimics, inhibitor and negative control (NC), CA12 overexpression plasmid, and small interfering RNA (siRNA) targeting CA12 and siRNA NC were provided by Guangzhou RiboBio Co., Ltd. and transfected into rat tracheal chondrocytes.

Techniques: Binding Assay, Mutagenesis, Luciferase, Reporter Assay, Transfection

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: CA12 suppressed rat tracheal chondrocyte proliferation and differentiation. Rat tracheal chondrocytes were transfected with CA12 overexpression plasmid or CA12 siRNA. (a) Transfection efficiency was determined by western blot. (b) Viability of rat tracheal chondrocytes assessed by CCK-8 assay. (c) Cell proliferation ability assessed by EdU staining. Red fluorescence: EdU; blue fluorescence: DAPI (scale bar = 50 μ m). (d) Cell apoptosis levels assessed by TUNEL staining (scale bar = 50 μ m). Red fluorescence: TUNEL; blue fluorescence: DAPI. (e) Cell apoptosis assessed by flow cytometry with Annexin V/propidium iodide staining. (f) Protein expression of E-cadherin and N-cadherin. (g) The effect of miR-548au-3p on differentiation of rat tracheal chondrocytes assessed with Alcian blue staining (scale bar = 100 μ m). (h) Aggrecan, Col2A1, MMP13, and ADAMTS4 protein levels detected by western blot. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, compared with vector or siRNA NC.

Article Snippet: The miR-548au-3p mimics, inhibitor and negative control (NC), CA12 overexpression plasmid, and small interfering RNA (siRNA) targeting CA12 and siRNA NC were provided by Guangzhou RiboBio Co., Ltd. and transfected into rat tracheal chondrocytes.

Techniques: Transfection, Over Expression, Plasmid Preparation, Western Blot, CCK-8 Assay, Staining, Fluorescence, TUNEL Assay, Flow Cytometry, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: The Role of the miR-548au-3p/CA12 Axis in Tracheal Chondrogenesis in Congenital Pulmonary Airway Malformations

doi: 10.1155/2023/6428579

Figure Lengend Snippet: Knockdown of CA12 abolished miR-548au-3p inhibitor mediated effects on rat tracheal chondrocytes. Rat tracheal chondrocytes were cotransfected with CA12 siRNA and miR-548au-3p inhibitor for 48 h. (a) Transfection efficiency was determined by western blot. (b) Viability of rat tracheal chondrocytes assessed by CCK-8 assay. (c) Cell proliferation ability assessed by EdU staining. Red fluorescence: EdU; blue fluorescence: DAPI (scale bar = 50 μ m). (d) Cell apoptosis assessed by TUNEL staining (scale bar = 50 μ m). Red fluorescence: TUNEL; blue fluorescence: DAPI. (e) Cell apoptosis assessed by flow cytometry with Annexin V/propidium iodide staining. (f) E-cadherin and N-cadherin protein levels assessed by western blot. (g) The effect of miR-548au-3p on chondrogenic differentiation of rat tracheal chondrocytes assessed by Alcian blue staining (scale bar = 100 μ m). (h) Aggrecan, Col2A1, MMP13, and ADAMTS4 protein levels assessed by western blot. ∗∗ p < 0.01 and ∗∗∗ p < 0.001, compared with NC or inhibitor siRNA NC.

Article Snippet: The miR-548au-3p mimics, inhibitor and negative control (NC), CA12 overexpression plasmid, and small interfering RNA (siRNA) targeting CA12 and siRNA NC were provided by Guangzhou RiboBio Co., Ltd. and transfected into rat tracheal chondrocytes.

Techniques: Knockdown, Transfection, Western Blot, CCK-8 Assay, Staining, Fluorescence, TUNEL Assay, Flow Cytometry