The PLA reagents were obtained from Millipore-Sigma (DUO92101-1KT). The assays were performed following the manufacturer’s protocol. The antibodies against SREBP2 (Cayman Chemical Company, 10007663), SREBP1 (ProteinTech, 14088-1-AP), DAXX (5G11 hybridoma supernatant) were used for the PLA and IF experiments. IF experiments were performed as described previously, and images were acquired with a Zeiss AxioPhot microscope equipped with an Exi Blue camera (Qimaging)86 (link).
Mahmud I., Tian G., Wang J., Hutchinson T.E., Kim B.J., Awasthee N., Hale S., Meng C., Moore A., Zhao L., Lewis J.E., Waddell A., Wu S., Steger J.M., Lydon M.L., Chait A., Zhao L.Y., Ding H., Li J.L., Purayil H.T., Huo Z., Daaka Y., Garrett T.J, & Liao D. (2023). DAXX drives de novo lipogenesis and contributes to tumorigenesis. Nature Communications, 14, 1927.
PLA reagents obtained from Millipore-Sigma (DUO92101-1KT)
dependent variables
Outcomes measured in PLA and IF experiments
control variables
Antibodies against SREBP2 (Cayman Chemical Company, 10007663), SREBP1 (ProteinTech, 14088-1-AP), and DAXX (5G11 hybridoma supernatant) used for PLA and IF experiments
IF experiments performed as described previously
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
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