Fig. 1

a Pellino-1 expression was detected by western blotting in tumor and adjacent non-tumor tissues of human breast cancer (normal n = 9, luminal n = 9, HER2⁺n = 9, and TNBC n = 9). b Kaplan–Meier plot showed overall survival of TNBC patients in tissue microarray sections (HBreD140Su03) depending on the expression of Pellino-1 (n = 100). c The strategy for studying the effects of Pellino-1 in patient-derived tumor xenograft (PDX) model from TNBC patients (n = 6 per group) and statistical analyses of the tumor weights in the PDX model after Pellino-1 knockdown. d Immunoblots showing the expression of SNAIL and SLUG after Pellino-1 knockdown (n = 3). e The indicated concentrations of Resistomycin were passed over immobilized Pellino-1-GST on CM5 sensor chips and the kinetic interaction of Resistomycin with Pellino-1 was determined with SPR analyses (n = 3). f Heat map presenting the expression levels of SNAIL and SLUG in 4 TNBC cell lines and 1 primary human TNBC cells after indicated concentrations of Resistomycin treatment (n = 3). g Primary TNBC cells and MDA-MB-231 cells were treated with Resistomycin for 24 h, and the transwell assay was applied to assess invasion (n = 3). The number of invading cells was calculated in three different fields. Scale bar, 20 μm. h MDA-MB-231 cells were injected into the mammary fat pad of nude mice unilaterally. One week later, mice were treated with Vehicle or Resistomycin, and infected with Ad-GFP or Ad-PELI1 for 3 weeks (n = 6 per group). Data are bioluminescence imaging of mice (top) and statistical analyses of lung metastatic nodules (down). i The binding mode of Resistomycin to the homology model of the FHA domain by using crystal structure of Pellino-2 (PDB code 3EGA) as the template. Pellino-2 is shown in ribbon plot representation. Compounds are labeled in color by atoms. The hydrogen bonds are labeled as dashed lines. The key amino acid residues for the binding are labeled as sticks. j SPR analyses of the kinetic interaction of Resistomycin and Pellino-1 F137A (n = 3). k The strategy for studying the anti-tumor effects of Resistomycin in the presence or absence of Pellino-1 or Pellino-1-F137A overexpression in vivo. l, m The indicated MDA-MB-231 cells were injected into the mammary fat pad of nude mice unilaterally. One week later, mice were treated with Vehicle or Resistomycin for 3 weeks (n = 5 per group). Data are statistical analyses of the tumor weights (l), macroscopic and histopathologic images of lung metastatic nodules and statistical analyses of these nodules (m). Scale bar, 200 μm. n Schematic diagram illustrates the role of Resistomycin in inhibition of TNBC progression through promoting SNAIL/SLUG degradation via binding with Pellino-1. *P < 0.05, **P < 0.01, ***P < 0.001