Fig. 5: CyMA-d disrupt posttranslational modifications and cytokine secretion and lead to cell death.

a Metabolic labeling scheme for protein palmitoylation or O-GlcNAcylation or sialylation. b Protein modification analysis in HeLa treated with 3a (12 h) via metabolic labeling (n = 10 for palmitoylation analysis, and n = 7 for O-GlcNAcylation or sialylation analysis). c CLSM of protein sialylation in HeLa cells treated with 3a (12 h). d Volcano plot of regulated palmitoylated proteins in KPCA-C cells treated with 3a (2 μM, 6 h) (n = 3). e Relocation of representative RTKs, GPCR and G-proteins in different cell lines with/without 3a (2 μM, 6 h). f, g ELISA of TGF-β1 (f) and VEGF (g) in conditioned media from indicated cells treated with 3a (500 nM) (n = 3). h GI₅₀ and GI₉₀ of 3a against various cell lines. i GI₅₀ and GI₉₀ of 3a and 6 in CES-overexpressing cells (HepG2, RAW264.7) ± BNPP (50 μM, 24 h). j HeLa viability after 3a treatment (24 h), with/without 2-BP (10 μM) or triacsin C (10 μM) (n = 3). k Viability of stimulated/non-stimulated KPCA-C cells treated with 3a for 24 hours. l Viability of KPCA-C cell spheroids treated with 3a for 72 hours (n = 3). m Live/dead staining of PDOTS treated with 3a ± immune checkpoint inhibitors (200 μg/mL anti-PD-1 and anti-PD-L1) (n = 3). n Body weight change of mice injected with vehicle or 3a (125 μg/mouse, three times a week for a total 8 injections) intraperitoneally in PBS (n = 4). o Number of metastases, (p) Tumor weight and (q) presence of ascites in KPCA-B bearing mice treated with vehicle, 3a alone (3.6 μg/mouse), ICB alone (50 μg/mouse anti-PD-L1 and 50 μg/mouse anti-CTLA-4), or combination (n = 5 for vehicle and combination, n = 4 for the rest). Scale bar = 20 μm. Data are mean ± s.d. (n-values as indicated in the panel). Statistical significance was determined by a two-tailed Student’s t test. Reproducibility and statistical details are provided in the Methods. Source data are provided as a Source Data file.