To characterize the global landscape of lysine lactylation (Kla) in Th17 cells, we cultured naïve CD4⁺ T cells isolated from Trmt61a-cKO and WT mice under Th17-polarizing conditions for 96 hours. At least 1 × 10⁷ cells per sample were harvested for proteomic analysis. Following tryptic digestion, Kla-modified peptides were enriched using anti-Kla antibodies and analyzed by LC–MS/MS.