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The cells are first treated with the specific inhibitor as described in the text before the addition of murine rIL-21. For the combinational treatment, BX795 is added at the beginning. This reagent acts as a potent B cell differentiation agent in that it bypasses the requirement for B cell receptor signal transduction for its activity. Cells are then cultured for another 48 h before harvest. For the experiments in BX795 without murine rIL-21, the cells are firstly cultured with BX795 before the addition of murine rIL-21. If not specified, unstimulated cells were harvested for the purpose of comparison. Cells were collected by centrifugation, washed in PBS once, then stained using a combination of CD11c, CD45, B220, and CD86 or CD80 antibodies (eBioscience). Surface molecules were stained with fluorescent conjugated antibodies (eBioscience). Single cell suspensions were first stained with CD11c, B220, and CD45. The cells were then washed with PBS and incubated with CD11c, CD45, B220, and CD86 or CD80 antibodies for 30 min at 4°C. For the definition of macrophage phenotypes, cells are stained with an anti-CD11c, anti-F4/80, anti-CD86, or anti-CD80 antibodies. usp 38 pdf free download

PTS1 KO mice are backcrossed onto BALB/c mice for more than 10 generations. We used the same KO mice for the different experimental studies and we made the same effort to perform experiments with these KO mice at the same time. Mice were housed and maintained under SPF conditions.

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The use of patient samples was approved by the Ethics Committee of Guizhou Medical University (Approval no. 2018-123-01) and informed consent was obtained from the patients. All animals were housed and maintained in specific pathogen-free conditions according to the recommendation of Guide for the Care and Use of Laboratory Animals of the National Institutes of Health with strict accordance with protocols approved by the Ethics Committee of Guizhou Medical University (Approval no.1900628)
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