Figure 2

Downregulation of MYO5B on mRNA and protein levels in human CRC. (A) MYO5B mRNA levels in bulk tissue of matched tumour (CRC, n=50) and non-tumour (normal, n=50) samples from CRC patients. Data are presented as mean±s.d. (***P<0.001, paired t-test). (B) MYO5B mRNA levels in laser-microdissected tumour (CRC, n=67) and non-tumour (normal, n=21) samples. Data are presented as mean±s.d. (***P<0.001, unpaired t-test). (C) In 20 out of 21 paired microdissected tumour samples, MYO5B mRNA levels were higher in non-tumour tissue compared to the matched tumour sample (***P<0.001, paired t-test). (D) Immunohistochemical staining of 74 paired TMAs of CRC patients. Left panel: representative image of MYO5B immunostaining on a section that contains normal (bottom) and cancerous tissue (top). Staining was mainly cytoplasmic and showed the expected accumulation of MYO5B at the apical brush border of normal enterocytes, as well as in tumoural cells in which differentiation was still preserved. Scale bar corresponds to 200 μm. Right panel: quantification of MYO5B staining of matched tumour (CRC; n=74) and non-tumour (normal; n=74) samples scored according to the staining intensity of MYO5B-positive cells (0=no signal, 1=mild signal, 2=moderate signal, and 3=strong signal). Two sections of cancerous as well as normal counterpart tissue were scored per patient and the mean value was considered. Data are presented as mean±s.d. (***P<0.001, paired t-test). (E) Representative pictures (left panel) and quantification (right panel) of immunohistochemical staining of MYO5B in patient samples of different TNM stages. Data are presented as mean±s.d. (**P<0.01, ***P<0.001, unpaired t-test). Scale bar corresponds to 200 μm. (F) MYO5B scoring of TMA samples after classification according to their histological grade. The information was missing for one patient. Data are presented as mean±s.d.; unpaired t-test. *P<0.05, **P<0.01, and ***P<0.001.