The construction of transcriptional regulatory networks of transcription factors (TFs) has become more important and attractive to understand the alterations of binding protein-dependent transcriptional activity that governs the changes in spatiotemporal expression of TF target genes required in various cellular processes. Therefore, identification of new inner modules including target genes and protein interactions involved in unveiled TF-based transcription networks is currently in the research spotlight. In this study, we reveal a possible SOX4-centered transcriptional network by the identification of novel binding partners and target genes of the TF SOX4 using various screening techniques. Lamin B2, barrier to autointegration factor 1, and apolipoprotein C-III were identified as novel interacting partners of SOX4 by yeast two-hybrid screening, and the genes encoding lysosomal-associated membrane protein 1, ubiquitin-conjugating enzyme E2S, and Map2k2 were identified as putative target genes of SOX4. Differently from the computational networks of TFs, we revealed a SOX4-centered physical network during myoblast differentiation. These results will provide opportunities to better understand the SOX4-centered transcriptional regulation network and TF-based specific gene expression in various cellular environments.