Fig. 3. PFKFB3 inhibition promotes tumour vessel normalisation and inhibits trastuzumab resistance.

a Representative immunofluorescence (IF) images of CD31 on tumours of trastuzumab-sensitive (Tra-S) and resistant (Tra-R) patients. Scale bar, 50 μm (×20). b Scanning electron microscopy (SEM) images of tumour vessels from trastuzumab-sensitive (Tra-S) and resistant (Tra-R) patients. c IF images and comparisons of VE-cadherin distribution, collagen type IV⁺ basement membrane (BM) coverage and αSMA⁺ pericyte coverage were presented as a percentage of length that lied along the CD31⁺ vascular endothelium in trastuzumab-sensitive (Tra-S) and resistant (Tra-R) patients. Scale bar, 50 μm (×20). Each experiment was performed at least triplicately and all statistical results are shown as mean ± SEM, based on Student’s t test, ***P < 0.001. d IF images and comparisons of VE-cadherin distribution, collagen type IV⁺ basement membrane (BM) coverage and αSMA⁺ pericyte coverage were presented as a percentage of length lying along the CD31⁺ vascular endothelium in subcutaneously tumours. Scale bar, 50 μm (×20). Values are mean ± SEM. *P < 0.05, **P < 0.01, ****P < 0.0001. e Images and comparisons of lectin-perfused and dextran-leaky tumour vessels. Scale bar, 50 μm (×20). Values are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. f Images of Hypoxyprobe⁺ areas in subcutaneous tumours. Hypoxyprobe was injected intraperitoneally 60 min before tumour sampling. Scale bar, 50 μm (×20). g Phalloidin staining revealing the comparable actin cytoskeleton of EC cultured with TRs knockdown PFKFB3 or NC. Scale bar, 5 μm (×200).