Extended Data Fig. 7: Manual FlowJo analysis confirms increased p-CREB + MFI and frequency in innate and adaptive immune cells. | Nature Immunology

Extended Data Fig. 7: Manual FlowJo analysis confirms increased p-CREB + MFI and frequency in innate and adaptive immune cells.

From: The transcription factor CREB1 is a mechanistic driver of immunogenicity and reduced HIV-1 acquisition following ALVAC vaccination

Extended Data Fig. 7

a, p-CREB signal was quantified following treatment with Forskolin (n = 3), a known inducer of cAMP signaling, showing that p-CREB induction results in a shift in signal but is not bi-modal. b, Gating strategy for analysis. c, Histogram of p-CREB1 MFI in all cells and Clusters 5, 16, 17 and 20 showing higher p-CREB1 in these clusters compared to all cell. d, Density plots of UMAP analysis from Fig. 4 show that cGAMP induced p-CREB + populations are returned to basal levels after 60 minutes which is suggestive of true phospho-signaling. e, Representative histograms showing shift in p-CREB activity in total CD4 + T cells following 1 μM cGAMP stimulation for 15 m. f, Manual flow analysis of cGAMP treated samples (n = 5) in Fig. 4 confirms significant increases in p-CREB MFI for total CD4 + T cells, B cells, NK cells and other innate cells. Statistical analysis performed using a two-sided paired t-test. Line represents median and error bars are standard error of the mean.

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