Fig. 2: Confocal microscopy of CPMV and RNA in macrophages.
From: Cellular fate of a plant virus immunotherapy candidate
a 3-D spatial resolution of CPMV RNAs and coat proteins colocalized with lysosomal associated membrane protein 1 (LAMP-1). Multiplexed immunofluorescence and RNA fluorescent in situ hybridization (FISH) detect CPMV coat proteins (green), RNA-1, and RNA-2 (white) colocalized with LAMP-1 (red). Scale bar to 100px. b Confocal images showing DAPI, LAMP-1, CPMV, and RNA channels at time points 0 and 24 h (which is 0 and 24 h additional incubation after CPMV was incubated with RAW 264.7 macrophage cells for 24 h). Fluorescent intensity of coat protein and RNAs decreases within 24 h, indicating degradation. Sustained High Mander’s Coefficient with RNA and LAMP-1 over time. At 0 h, Mander’s Coefficient with CPMV to RNA-1 and RNA-2 are 0.934 and 0.901, respectively. At 24 h, Mander’s Coefficient with CPMV to RNA-1 and RNA-2 are 0.847 and 0.878, respectively. Images taken at 60× with oil objective. Scale bar to 10 μM.
