High mobility group box 1 (HMGB1) is an alarmin that initiates and regulates inflammation and immune responses in several ways, such as inducing cytokine release, cell migration and cell differentiation. During homeostatic conditions, HMGB1 is mainly located in the nuclei where it binds DNA and regulates DNA repair, recombination and transcription. However, cytosolic HMGB1 has been reported to be involved in autophagy and cell survival. Our unpublished proximity proteomics data further indicates other processes that HMGB1 is involved in. To investigate the role of intracellular HMGB1, we have RNA-sequenced resting and LPS-stressed wildtype (n=3) and HMGB1 knockout fibroblasts (n=3). To analyze the sequencing data, we aim to use UPPMAX (STAR, HISAT) to convert the fastQ (43 GB) files into SAM/BAM files. Thereafter, differential gene expression will be determined and genes of interest will be validated with qPCR. In the longer run, we aim to reveal the molecular role of HMGB1 in previously undescribed mechanisms.