Figure 7. P4HA2‐mediated collagen synthesis attenuates cancer cell differentiation.

1. Western blot analyses of expression levels of TTF‐1 and Mucin 5B in H1299^control and H1299^RASSF1A cells with band intensity quantification ratio determine by ImageJ analyser.
2. Representative images of H&E and immunohistochemical staining for lung differentiation markers TTF‐1 and Mucin 5B for at least two independent regions of n = 4 H1299^control and n = 2 H1299^RASSF1A primary tumours at day 17 with quantification (right) of TTF‐1 based on nuclear intensity. Scale bars: 100 μm. Statistical significance was determined by Student's t‐test (two‐tailed). Error bars represent the mean ± SEM
3. Representative images of immunofluorescence staining of spheroids grown in collagen matrix (2 mg/ml) for differentiation markers TTF‐1 (Green) and Mucin 5B (Red). Scale bars: 50 μm; Zoom: 10 μm.
4. Bright‐field images of H1299^control and H1299^RASSF1A spheroid differentiation on Matrigel matrix after 24 h when 3D spheroids were seeded on three‐dimensional substrate. Scale bars: 200 μm.

Source data are available online for this figure.