Figure 3.

Inhibiting LGR5 glycosylation results in reduced localisation within the cis-Golgi network. Confocal laser scanning microscopy Z-sections showing intracellular LGR5 expression in fixed and permeabilised (A) LoVo and (B) LS174t-LGR5 cells following 24 h treatment with DMSO control or 1 μg ml⁻¹ TMC. Merged (LGR5 and Grasp65 only), LGR5 (green), Grasp65 (G65; magenta), E-cadherin (E-Cad; red) and DAPI (blue) images are shown. Areas of co-localisation between LGR5 and Grasp65 are indicated by resulting white colour (highlighted by arrowheads). Overall correlation between LGR5 and Grasp65 signal in DMSO control or TMC treated (C) LoVo and (D) LS174t-LGR5 cells, as determined by thresholded Pearsons Correlation Coefficient (−1= inverse correlation, 0= no correlation, +1= positive correlation). Proportion of Grasp65 signal co-localised with LGR5 fluorescence in DMSO control or TMC treated cells is also shown as determined by the Mander's Colocalisation Coefficient. Data represent mean ±1 s.d. as deduced from a minimum of 20 random fields, n=3. Statistical significance is denoted by ***P<0.001, as analysed by students t-test.