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. Author manuscript; available in PMC: 2012 Jul 1.
Published in final edited form as: Biochim Biophys Acta. 2011 Jun 13;1809(7):306–315. doi: 10.1016/j.bbagrm.2011.05.016

Fig. 2. Expression of mRNA and protein in SW480 after histidinol treatment.

Fig. 2

a) mRNA level was quantified in human colon cancer cell line SW480 after histidinol (HisOH) treatment. SW480 cells were grown in MEM, MEM without amino acid, and MEM containing 2, 4, 8, or 16 μmol/L histidinol for 8 h. Relative mRNA level was determined by quantitative real time RT-PCR. Data were normalized by the internal control L7a. b) A representative blot from Western blotting analysis for ATF4, p-eIF2α and p-ERK protein expression. c) Protein level was quantified in SW480 by Western blotting with 16 mmol/L HisOH treatment. A non-specific band was used as the loading control to normalize the raw data. Each bar represents the mean ± S. D. of three independent experiments. Asterisks (*) represent statistical significance compared to values in MEM (P < 0.05). d) The MEM-, MEM-AA-, and MEM+HisOH-treated cells were fixed on glass cover slips and subsequently used for immunofluorescent staining. DKK1 protein was analyzed after immunofluorescent staining using an antibody against DKK1 protein and an Alexa Fluor 647-labeled secondary antibody (red). The cover slips were also counterstained with Hoechst 33342 fluorescent staining for nucleus (blue). The two pictures were overlaid to show DKK1 staining on top of the nuclear staining (merged). Cells treated with in vitro transfection of DKK1-containing plasmid was used as a positive control for the staining of DKK1 protein.