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Proteins, Expression, Isolation and Analysis | An equal number of cells were collected and subjected to immunoblot analysis to determine protein expression levels of Bcl-6 (BD Biosciences, 561520, dilution 1:500), Blimp-1 (Genscript, A01647, dilution 1:500), STAT5 (Santa Cruz, sc-835, dilution 1:5,000), phospho-STAT5 (pY694, BD Biosciences, 611964, dilution 1:5,000) and V5-tagged proteins (Invitrogen, R960-25, dilution 1:5,000).... GAPDH (Santa Cruz, sc-25778, dilution 1:2,500) or b-actin (Genscript, A00730, dilution 1:10,000) expression was monitored to ensure equal protein loading. | Get A Quote |
The transcriptional repressor Bcl-6 is linked to the development of both CD4(+) T follicular helper (TFH) and central memory T (TCM) cells. Here, we demonstrate that in response to decreased IL-2 signalling, T helper 1 (TH1) cells upregulate Bcl-6 and co-initiate TFH- and TCM-like gene programs, including expression of the cytokine receptors IL-6Rα and IL-7R. Exposure of this potentially bi-potent cell population to IL-6 favours the TFH gene program, whereas IL-7 signalling represses TFH-associated genes including Bcl6 and Cxcr5, but not the TCM-related genes Klf2 and Sell. Mechanistically, IL-7-dependent activation of STAT5 contributes to Bcl-6 repression. Importantly, antigen-specific IL-6Rα(+)IL-7R(+) CD4(... More