Quantifying Oxidized LDL Uptake in U937 Macrophages

The U937 macrophages were incubated in the presence of 10 μg/mL Dil-ox-LDL in RPMI 1640 medium supplemented with 10% fetal bovine serum including 100 μg/mL streptomycin and 100 U/mL penicillin at 37 °C in 5% CO₂ for 18 h [21 (link),26 (link)]. After gentle washing with PBS three times, the adherent cells were mounted in Vectashield mounting medium (H-1500, Vector Laboratories, Burlingame, CA, USA) and were imaged with BZ-X710 microscope/software (Keyence, Osaka, Japan). The fluorescent intensity of red color area per cells was quantified as described previously [21 (link),26 (link)].

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Yashima H., Terasaki M., Sotokawauchi A., Matsui T., Mori Y., Saito T., Osaka N., Kushima H., Hiromura M., Ohara M., Fukui T, & Yamagishi S.I. (2020). AGE-RAGE Axis Stimulates Oxidized LDL Uptake into Macrophages through Cyclin-Dependent Kinase 5-CD36 Pathway via Oxidative Stress Generation. International Journal of Molecular Sciences, 21(23), 9263.

Publication 2020

Vectashield mounting medium (H-1500, BZ-X710 microscope/software

Cells Fetal bovine serum Macrophages Microscope Ox ldl Penicillin Streptomycin Vector

Corresponding Organization : Showa University

Other organizations : Kurume University

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Variable analysis

independent variables

Concentration of Dil-ox-LDL (10 μg/mL)

dependent variables

Fluorescent intensity of red color area per cells

control variables

Incubation medium (RPMI 1640 with 10% fetal bovine serum, 100 μg/mL streptomycin, and 100 U/mL penicillin)
Incubation conditions (37 °C, 5% CO2, 18 h)

controls

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

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