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. 2015 Jun 25;10(6):e0131453. doi: 10.1371/journal.pone.0131453

Fig 1. BMP signaling is activated by TCR stimulation in naive CD4+ T cells.

Fig 1

Freshly isolated human peripheral blood naive CD4+ T cells were stimulated with anti-CD3/CD28 mAb. (A) Transcripts for several components of the canonical BMP signaling pathway were determined by real-time PCR ex vivo (0h) or after 6 days of stimulation (6d). GNB2L1 was used as endogenous control. Means ± SD of at least three independent experiments run in duplicates are shown. Note the logarithmic scale on y-axis. (B) Percentage of BMPRIA+ cells detected by flow cytometry throughout the culture. Bars represent the mean ± SD of two to five independent experiments. (C) Expression of BMPRIA and CD25 in T cells (upper dot plots) and differential expression of BMPRIA in the CD25- and CD25+ cell populations (lower histograms) during activation. A representative experiment out of four is shown. (D) Expression of BMPRIA in T cells cultured with different stimuli. Grey histograms represent isotype controls. Similar stainings were obtained in two to three independent experiments. mDCs: mature dendritic cells; PHA: Phytohaemagglutinin; Ck: cytokine cocktail (rhIL-2, rhIL-6, rhTNF-α) (E) Determination of BMP2/4 and BMP6 production by flow cytometry in T cells cultured in media alone (grey histograms) or in the presence of anti-CD3/CD28 mAb (black histograms). Grey filled histograms represent isotype control stainings. A representative experiment out of three is shown. (F) Expression of CD25 and phosphorylated BR-Smad (pBR-Smads) during activation. For comparison, T cells were kept in culture media alone. Results are representative of three independent experiments.