Immunoblotting Analysis of Cellular Proteins

Immunoblotting was performed as described previously (43 (link)). Briefly, protein samples were subjected to SDS-PAGE and transferred to a polyvinylidene difluoride membrane probed with the following antibodies: Drp1 (BD Biosciences, 610296), CD144 (Invitrogen, 14-1441-81), p–Drp1 Ser637 (Cell Signaling Technology, 6319), β-actin (Sigma-Aldrich, A1978), VCAM-1 (Santa Cruz Biotechnology Inc., sc-13160), CD31 (MilliporeSigma, MAB1398Z), PDK1 (Santa Cruz Biotechnology Inc., sc-515944), HK2 (Santa Cruz Biotechnology Inc., sc-374091), T-eNOS (BD Biosciences, 610296), OPA1 (BD Biosciences, 612606), Mfn2 (Santa Cruz Biotechnology Inc., sc-100560), Fis1 (Sigma-Aldrich, HPA017430), and α-tubulin (Sigma-Aldrich, T9026). Detailed methods are provided in the Supplemental Methods.

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Hong S.G., Shin J., Choi S.Y., Powers J.C., Meister B.M., Sayoc J., Son J.S., Tierney R., Recchia F.A., Brown M.D., Yang X, & Park J.Y. (2022). Flow pattern–dependent mitochondrial dynamics regulates the metabolic profile and inflammatory state of endothelial cells. JCI Insight, 7(18), e159286.

Publication 2022

CD144 p–Drp1 Ser637 β-actin VCAM-1 T-eNOS α-tubulin

Actin Antibodies Enos Membrane Mfn2 Opa1 Pdk1 Polyvinylidene difluoride Protein Sds page Vcam 1 α tubulin

Corresponding Organization : Baylor University

Other organizations : University of Maryland, Baltimore, Istituto di Fisiologia Clinica, Scuola Superiore Sant'Anna, National Research Council, University of Maryland, College Park

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Variable analysis

independent variables

Antibodies used for immunoblotting: Drp1, CD144, p-Drp1 Ser637, VCAM-1, CD31, PDK1, HK2, T-eNOS, OPA1, Mfn2, Fis1

dependent variables

Protein expression levels detected by immunoblotting

control variables

β-actin
α-tubulin

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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