Nature - USA (2019-07-18)

4

nature research | reporting summary

October 2018

Purity of magnetic bead CD4+ T cell isolation was determined by flow cytometry with >90% CD4 expressing cells used for all

experiments.

Gating strategy For all experiments measuring Ifng-Kat reporter expression, cells were gated on FSC-A high, DAPI low >> lymphocyte FSC-A, SSC-
a >> CD4 positive, GFP positive based on clear distinctions between populations in the pseudo-color plot. Cells were binned into
4 quartiles of katushka expression. This gating strategy is shown in Extended Data Figure 4a.

For experiments measuring intracellular cytokine in retrovirally transduced cells, gating was as followed: FSC-A, SSC-A live cells

>> SSC-W, SSC-H singlets >> FSC-H, FSC-W singlets >> CD4 positive, GFP positive live transduced cells.

For experiments measuring intracellular cytokine in cells not transduced, gating was as followed: FSC-A, SSC-A live cells >> SSC-

W, SSC-H singlets >> FSC-H, FSC-W singlets >> CD4 positive, LIVE/DEAD Negative.

Tick this box to confirm that a figure exemplifying the gating strategy is provided in the Supplementary Information.