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. 2011 Jun 17;39(16):e108. doi: 10.1093/nar/gkr465

Figure 7.

Figure 7.

Detection of full-length acGFP restoration by western blot analysis. After co-transfection of target molecule and either RTM 2 (lane 6), 4 (lane 7), or 3 (lane 4), acGFP protein (27 kDa) was detected by western blot analysis. As positive control, total-cell extract from acGFP control vector (acGFP cloned in pcDNA 3.1D/V5-HIS vector, Invitrogen) transfected HEK293AD cells was also included (lane 2). Lane 3 demonstrates a significant increase (∼3-fold) in the amount of acGFP protein in acGFP positive cells collected by FACS sorting of HEK293AD cells co-transfected with RTM 2 and target expression plasmids (lane 3) compared to total cell extract from co-transfected HEK293AD cells without sorting (lane 6). No detectable acGFP expression was visible after single transfection of target molecule (lane 11), RTM 4 (lane 10) or RTM 2 (lane 9) into HEK293AD cells. No expression of acGFP was detected by western blot analysis of HEK293AD cells co-transfected with target molecule and either RTM 1 (lane 5) or RTM 0 (which is missing a specific BD for intron 52, lane 8). Annexin I (37 kDa) was included as the loading control for this experiment. Lane 1: Precision Plus Protein WesternC Standards (Biorad).