Purpose : Transcription factor expression in cell(Raw264.7)

Method :

1.(ver.1) pCIneo constitutive expression vector + target cDNA are inserted, (ver2.Flag tag added to 3`end of target cDNA), my target gene cDNA was inserted to pCIneo plasmid (Nhe1, Not1 RE used)

After sequencing, midi-prep, plasmid transfected to Raw264.7 cell(reagent FugeneHD used)

Result : qRT-PCR result : after 12h, target mRNA expression increased

Western blot : not detected , used antibody : target protein antibody, FLAG tag antibody

when all above are considered, i think that the problem is translation step.

and the protein that i try to express is not expressed at macrophage , orignially the protein expressed in neuronal cell

What should i consider to solve this problem?