Cancer-associated fibroblasts (CAFs) are a critical heterogeneous component of the tumor microenvironment (TME), playing dual roles in tumor progression. They contribute to immune evasion, epithelial-to-mesenchymal transition (EMT), angiogenesis, and extracellular matrix remodeling, while certain subsets of CAFs exhibit tumor-suppressive functions. Understanding the diversity and plasticity of CAFs is essential for developing effective therapeutic strategies. Identifying key regulators of CAFs would expand the possibility of CAF-targeted cancer therapy. Here, we present step-by-step experimental methodologies to explore potential therapeutic targets aimed at selectively suppressing the pro-tumoral activities of CAFs, while preserving or enhancing their tumor-suppressive functions.