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Polymicrobial infections are more challenging to treat and are recognized as responsible for significant morbidity and mortality. It has been demonstrated that multiple Gram-negative organisms take advantage of the effects of <i>Staphylococcus aureus</i> α-toxin on mucosal host defense, resulting in proliferation and dissemination of the co-infecting pathogens. Through phenotypic approaches, we observed a decrease in the motility of <i>A. baumannii</i> A118 after exposure to cell-free conditioned media (CFCM) of <i>S. aureus</i> strains, USA300 and LS1. However, the motility of <i>A. baumannii</i> A118 was increased after exposure to the CFCM of <i>S. aureus</i> strains USA300 Δ<i>hla</i> and <i>S. aureus</i> LSI Δ<i>agrA</i>. Hemolytic activity was seen in A118, in the presence of CFCM of <i>S. aureus</i> LS1. Further, <i>A. baumannii</i> A118 showed an increase in biofilm formation and antibiotic resistance to tetracycline, in the presence of CFCM of <i>S. aureus</i> USA300. Transcriptomic analysis of <i>A. baumannii</i> A118, with the addition of CFCM from <i>S. aureus</i> USA300, was carried out to study <i>A. baumannii</i> response to <i>S. aureus’</i> released molecules. The RNA-seq data analysis showed a total of 463 differentially expressed genes, associated with a wide variety of functions, such as biofilm formation, virulence, and antibiotic susceptibility, among others. The present results showed that <i>A. baumannii</i> can sense and respond to molecules secreted by <i>S. aureus</i>. These findings demonstrate that <i>A. baumannii</i> may perceive and respond to changes in its environment; specifically, when in the presence of CFCM from <i>S. aureus</i>.