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. 2015 Aug 31;16(10):1394–1408. doi: 10.15252/embr.201540107

Figure EV3. Loss of PHD2 suppresses CAF‐induced matrix remodelling and invasion.

Figure EV3

  1. Bars show quantification of the longest distance of HN‐CAFs plated in 3D collagen I/Matrigels. Bars indicate mean ± s.d. *P < 0.05; **P < 0.01; ***P < 0.001; one‐way ANOVA test. n = 3 experimental repeats.
  2. F‐actin staining of V‐CAFs grown on top of collagen I/Matrigels. Bars show quantification of the longest distance of V‐CAFs. Bars indicate mean ± s.d. **P < 0.01; ***P < 0.001; one‐way ANOVA test. n = 2 experimental repeats.
  3. Validation of multiple siRNAs targeting PHD2 by Western blotting. Validation of multiple siRNAs targeting PHD1&3 by qPCR. Bars show mean ± s.d. n = 2 independent experimental repeats done in triplicate.
  4. Depletion of PHD2 decreases V‐CAF‐induced invasion. Bars show mean ± s.d. invasive index. n = 2 experimental repeats.
  5. Depletion of PHD2 using multiple siRNAs decreases HN‐CAF‐induced invasion. Bars show mean ± s.d. invasive index. n > 3 experimental repeats. *P < 0.05; **P < 0.01; ***P < 0.001; unpaired Student's t‐test (two‐tailed).
  6. PHD2 regulates αSMA mRNA levels. V‐CAFs were siRNA‐depleted for PHD2, and mRNA levels of αSMA were quantified by qPCR 72 h post‐transfection. CAFs were plated on gels. Each data point represents an independent experiment done in triplicate. Line and error bars indicate mean ± s.d.
  7. Immunoblotting analyses of the HIF‐1α, PHD2 and MMP14. siRNA‐depleted V‐CAFs were grown on gels for 72 h post‐transfection.
  8. Immunoblotting validation of the rescue experiment presented in Fig 5G.