Immunofluorescence Staining of Naïve CD4 T Cells

Naïve CD4 T cells were isolated and cultured as described above, followed by immunofluorescence staining using a method described previously^{6 (link)}. Rabbit anti-53BP1 (Cell Signaling) and mouse TRF1 (Thermo Fisher) were used as primary antibodies and anti-rabbit IgG-Alexa Fluor 488 and anti-mouse IgG- Alexa Fluor 555 (Invitrogen) were used as secondary antibodies. Then, cells were washed and mounted with DAPI Fluoromount-G (SouthernBiotech, Birmingham, AL). Images were acquired with a confocal laser-scanning inverted microscope (Leica Confocal, Model TCS sp8, Germany).

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Nguyen L.N., Zhao J., Cao D., Dang X., Wang L., Lian J., Zhang Y., Jia Z., Wu X.Y., Morrison Z., Xie Q., Ji Y., Zhang Z., El Gazzar M., Ning S., Moorman J.P, & Yao Z.Q. (2018). Inhibition of TRF2 accelerates telomere attrition and DNA damage in naïve CD4 T cells during HCV infection. Cell Death & Disease, 9(9), 900.

Publication 2018

Rabbit anti-53BP1 mouse TRF1 anti-rabbit IgG-Alexa Fluor 488 anti-mouse IgG- Alexa Fluor 555 DAPI Fluoromount-G Model TCS sp8

53bp1 Alexa fluor 488 Alexa fluor 555 Anti igg Antibodies Cd4 t cells Cells Dapi Immunofluorescence Microscope Mouse Rabbit

Corresponding Organization :

Other organizations : East Tennessee State University, Air Force Medical University, 302 Military Hospital of China

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Variable analysis

independent variables

Isolation and culturing of naïve CD4 T cells

dependent variables

Immunofluorescence staining of 53BP1 and TRF1 proteins
Microscopic imaging of the stained cells

control variables

The method for immunofluorescence staining described previously in reference 6

controls

Positive control: Not specified
Negative control: Not specified

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