CXCR5+ monocyte emigration impairs the radiation-induced antitumor immune response

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Raw-MC38 untreated IR3 IR7 Gene expression profiles were generated on a total of 6 tumor tissue samples. Tumor tissue derived from three or seven days after IR were compared to untreated tumor (each group n=2). Raw-monocytes macrophages CXCR5- and CXCR5+ monocytes were separated by fluorescence-activated cell sorting from 50% TS treated BM derived monocytes. To obtain CXCR5- and CXCR5+ macrophages，CXCR5+ monocytes were cultured in induce medium for 3 days. These two subpopulations were separated by fluorescence-activated cell sorting. Raw-tumor infiltrated mon The CXCR5- and CXCR5⁺ monocytes were sorted from irradiated tumors and M-MDSCs were (Ly6ChighLy6Gow) sorted from untreated tumor. Sequencing libraries were generated using Hieff NGS Ultima Dual-mode mRNA Library Prep Kit for Illumina (Yeasen Biotechnology (Shanghai) Co., Ltd.) following manufacturer’s recommendations and index codes were added to attribute sequences to each sample. Raw data (raw reads) of fastq format were firstly processed through in-house perl scripts. The StringTie Reference Annotation Based Transcript assembly method was used to construct and identify both known and novel transcripts from Hisat2 alignment results.

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