Immunoblotting Analysis of Protein Extracts

Mouse cells were lysed as described previously^{6 (link)}. Protein extracts were fractionated on 8% (for LMP1 immunoblot), 10% (for Trp1 immunoblot) or 15% (for Survivin and EphA2 immunoblots) sodium dodecyl sulfate polyacrylamide gels, transferred onto polyvinylidene difluoride membranes and immunoblotted with the following primary antibodies: anti-LMP1 (clone S12, 1:4; obtained as described previously^{9 (link)}), anti-Survivin (clone 71G4B7, 1:1000; Cell Signaling), anti-EphA2 (clone C-3, 1:200; Santa Cruz), anti-Trp1 (clone TA99, 1:100; Santa Cruz), anti-Flag (clone M2, 1:5000; Sigma) and anti-GAPDH (clone 2118, 1:1000; Cell Signaling). Images were taken with ImageQuant LAS 4000 using ImageQuant LAS 4000 v1.2 software (both GE Healthcare Life Sciences).

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Choi I.K., Wang Z., Ke Q., Hong M., Paul DW J.r., Fernandes S.M., Hu Z., Stevens J., Guleria I., Kim H.J., Cantor H., Wucherpfennig K.W., Brown J.R., Ritz J, & Zhang B. (2020). Mechanism of EBV inducing anti-tumour immunity and its therapeutic use. Nature, 590(7844), 157-162.

Publication 2020

anti-LMP1 (clone S12, anti-Survivin (clone 71G4B7, anti-EphA2 (clone C-3, anti-Trp1 (clone TA99, anti-Flag (clone M2, anti-GAPDH (clone 2118 ImageQuant LAS 4000 ImageQuant LAS 4000 v1.2 software

Antibodies Cells Clone Gapdh Membranes Mouse Polyacrylamide gels Polyvinylidene difluoride Protein Sodium dodecyl sulfate Survivin Trp1

Corresponding Organization :

Other organizations : Dana-Farber Cancer Institute, Brigham and Women's Hospital, Harvard University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of LMP1, Trp1, Survivin, and EphA2

control variables

None explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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