Immunofluorescence Staining of Cellular Proteins

Immunofluorescence was performed as previously described [17 , 20 (link)]. Briefly, after treatments, cells were fixed, permeabilized, and stained with primary antibodies against DHX9 (No.17721–1-AP, Proteintech), p65 (No.66535–1-Ig, Proteintech), p65 (phospho S536) (p-p65, sc-136548, Santa Cruz), F4/80 (No.28463–1-AP, Proteintech) at 4 °C overnight. After incubating with the second antibodies, cellular nuclei were stained with DAPI (0.5 μg/ml, Cell signaling) and then imaged by a Carl Zeiss LSM 800 confocal.

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Huangfu N., Ma H., Tian M., Zhang J., Wang Y., Li Z., Chen X, & Cui H. (2023). DHX9 Strengthens Atherosclerosis Progression By Promoting Inflammation in Macrophages. Inflammation, 46(5), 1725-1738.

Publication 2023

(No.66535–1-Ig, DAPI LSM 800

After treatments Antibodies Cells Cellular nuclei Dapi Immunofluorescence

Corresponding Organization :

Other organizations : Ningbo First Hospital, Ningbo University

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Variable analysis

independent variables

Treatments

dependent variables

Localization and expression of DHX9, p65, and phosphorylated p65 (p-p65)
Localization and expression of F4/80

control variables

Cellular nuclei stained with DAPI

controls

Positive control: Not specified.
Negative control: Not specified.

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