Figure 7. Influence of cell type on
promoter-directed reporter gene expression. Promoter constructs were
transfected into wMC (rat Müller cells; red), NIH/3T3 (mouse
fibroblasts; green), or Neuro-2a (mouse neuroblastoma; blue). All
values were normalized to pFUGW and plotted according to percent
enhanced green fluorescent protein (eGFP) positive cells (A) or
mean fluorescence intensities (MFI; B). A: Percent eGFP
expressing cells are plotted for control (pFUGW, pFTMGW, and VMR2) and
xperimental (GLUL 500 ± untranslated region (UTR), S100B
500±UTR, and RLBP1 500±UTR) constructs. Note that transfection
efficiency of Neuro-2a cells was significantly higher overall (95%
positive cells versus ~20% for wMC and NIH/3T3), which, consequently,
resulted in significantly higher expression in the pFTM3GW control.
Nevertheless, when normalized to pFUGW, cell type had little influence
on the relative percentage of cells that express eGFP, with the
exception of a doubling in the number of NIH/3T3 cells transfected with
the RLBP1 500+UTR construct. B: Though absolute eGFP expression
levels were quite different for the three cell types examined, similar
results to those in (A) were obtained when the MFIs were
normalized to pFUGW. Perhaps an overall trend is noticeable; NIH/3T3
cells (green) tended to offer slightly higher relative eGFP expression,
and Neuro-2a cells (blue) tended to exhibit slightly lower eGFP
expression than that observed for wMC Müller cells (red). No TFX
indicates no transfection. Error bars represent 1 standard deviation.