Fig. 5.

The 3′UTR GQ elements are required for FA-induced cFPGS mRNA transport to cell protrusions. A–P On day 12 of FA deprivation, HeLa cells were co-transfected with MCP-GFP and F-MS2 (A–J) or F-MS2-dmGQ (K–O). Each transfection was divided into 2 wells and left to recover. At day 14, control cells remained FA-free (A–E) and the rest were subjected to a 15-min FA pulse (F–O), prior to fixation and IF microscopy. Q On day 13 of FA-deprivation, HeLa cells were co-transfected with MCP-GFP and F-MS2 along with ASOs as indicated. The next day, cells were subjected to a 15-min FA pulse, prior to fixation and microscopy. The localization of cFPGS mRNA was indicated by MCP-GFP (green, 488 nm), FLAG-cFPGS protein was detected by an anti-FLAG antibody (red, 543 nm), and F-actin was stained by DyLight 650 Phalloidin (white, 630 nm). Cells were scanned and manually counted using a confocal microscope (×63 magnitude). The scale bars denote 10 μm. P, Q Percentage of cells displaying protrusion-localized cFPGS mRNA. Shown are the mean percentage of cells ± S.D. from three independent experiments, n = total number of cells counted. *p <0.0006, **p < 0.00003