Fig. 5
Multiplex detection in RNAscope V2 assay for simultaneous visualization of HvGAPDH and Rga1 mRNA on barley line Q21861 leaf sections. Confocal Image capturing on Zeiss LSM-700 without DIC (a, c) and with DIC (b, d) transmitted light detcector T-PMT. (a, b.) Simultaneous visualization of HvGAPDH (C1 channel probe hybridized with TSA plus® fluorescein, green fluorescence dots) and Rpg1 (C2 channel probe hybridized with TSA plus® Cyanine3, orange fluorescence dots). Majority of Rpg1 expression was found to be associated with subsidiary cells associated with barley guard cells in the epidermal layer. DAPI stain was used for nuclear staining. (c, d.) A 3plex negative control with DAPI staining, without any probe specific fluorescence