FIGURES AND TABLES
FROM:
Differential expression of DHHC9 in microsatellite stable and instable human colorectal cancer subgroups
F Mansilla, K Birkenkamp-Demtroder, M Kruhøffer, F B Sørensen, C L Andersen, P Laiho, L A Aaltonen, H W Verspaget and T F Ørntoft
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Figure 1.
Microarray analysis of 168 samples representing molecular subgroups of CRCs. Transcript expression level of (A) DHHC9 and (B) H-ras on the U133plus2.0 Gene Chip. The black bars correspond to CRC samples, the grey bars to normal mucosa. Expression values are given as log 2 values and all data are normalised. (C) MSS/MSI study. Median log 2 values and standard deviations of normal biopsies (median normal, n=10), CRC patients (median cancer, n=168), MSI (CRC patients with MSI, n=35), MSS (CRC patients with MSS, n=118).
Full figure and legend (75K)
Figure 2.
Immunohistochemical analysis of DHHC9 expression. (A) normal mucosa, (B) Dukes A adenocarcinoma adjacent to premalignant tissue, (C) Dukes A adenocarcinoma (
200), (D) Dukes D adenocarcinoma, (E) lymph node metastasis and (F) liver metastasis ( 400). DHHC9 protein expression in MSS vs MSI colon adenocarcinomas. Microsatellite stable tumours (G–L), MSI-H tumours (M–R) ( 200).
Figure 3.
Immunostaining and Western blotting of transiently transfected COS7 cells and immunofluorescence applied to an MSS adenocarcinoma. (A–C) Confocal microscope, (D) Zeiss Microscope: 100. (A) DHHC9, (B) endogenous 58K protein, (C) merged DHHC9 and 58K protein, (D) DHHC9 Golgi localisation detail. DAPI-stained nuclei are blue. (E) Western blotting of extracts from DHHC9-transfected COS7 cells (MW, molecular weight marker; 1, cells transfected with an empty vector; 2, cells transfected with wild-type DHHC9). (F–K) IF, (F) DHHC9, (G) 58K protein, (H) DAPI, (I) merge (F) and (G); (J) merge (F), (G) and (H); (K) DAB staining of the same tumour.
Figure 4.
Proliferation assay. Several different cell lines with MSS (SW480, CaCo2) or MSI (HCT15, HCT116, LS174 TR4, DLD-1) were transfected with DHHC9 or a mock empty vector. Forty-eight hours post-transfection cells were added to 1 dye binding solution, and fluorescence was measured. Only MSS cell lines SW480 and CaCo2 showed a significantly decreased proliferation rate. Interestingly, also DLD-1 cells, with the wnt pathway switched off, showed a similar decreased proliferation rate.
![Table 2 - Immunohistochemical analysis of DHHC9 protein expression applying a 1[thinsp]:[thinsp]250 dilution of anti-DHCC9 antibody on 34 CRC tissue samples - Unfortunately we are unable to provide accessible alternative text for this. If you require assistance to access this image, please contact help@nature.com or the author](/common/images/table_thumb.gif)
