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. Author manuscript; available in PMC: 2014 Oct 1.
Published in final edited form as: Am J Med Genet A. 2013 Aug 16;161(10):2519–2527. doi: 10.1002/ajmg.a.36128

Figure 3.

Figure 3

Functional analysis of patient’s mutations. Wild type and mutant FLAG-tagged constructs were transiently transfected into HEK-293 cells. A) Protein levels as assessed by western blotting against the FLAG tag. The approximate size of the bands is 37 kDa. A wild type construct with no FLAG tag was used as a negative control, and β-actin levels were used as a loading control. B) As a control, RT-PCR was used to confirm transcription of plasmid-encoded B4GALT7 sequence in the transfected cell lines. Lane 1 is a RT-PCR off of RNA isolated from untransfected HEK-293 cells. Lane 2 is a PCR done directly from a WT-FLAG plasmid, performed as a positive control. Lanes 3, 4, 5, and 6 are RT-PCRs from RNA isolated from cells transfected with WT, WT-FLAG, L41P-FLAG, and R270C-FLAG plasmids, respectively.