Fig. 4

Immunological analyses of stems harvested from wild-type and fpgs1-1 plants. a Glycome profiling of sequential extracts prepared from the cell walls isolated from 35-day-old stems of fpgs1-1 and wild-type plants. The data are the average of three independent biological replicates. The various extraction reagents used are indicated at the bottom of the figure. The bar graphs above show amounts of carbohydrate materials released at each extraction step. Please note that these are sequential extracts (and not individual treatments) and reduced amounts released in last three extracts in mutant lines are potentially caused by the loss of excess carbohydrate materials in earlier lesser harsh extraction steps such as oxalate extraction. The panel on the right depicts the clades of monoclonal antibodies that recognize most major classes of plant cell wall glycans. The dotted boxes and arrows highlight the differences in the glycome profiles between fpgs1-1 and wild-type plants. The yellow-black scale indicates the strength of the ELISA signal: bright yellow color depicts the strongest binding and the black color indicates no binding. b Immunofluorescence labeling with four selected antibodies representing different clades of glycans. Tissue was harvested from 35-day-old plants and fixed and sectioned as described in “Methods”. (Scale bar 25 µm and is applicable to all images)