To examine
the TMV coat protein (cp) and tobacco ubiquitination levels, proteins
were extracted from tobacco seedlings that had been treated with PBS
and TMV solution. Samples were collected at different time points.
We performed the western blot analysis using a commercially available
antibody against ubiquitination as previously described.32 (link),63 (link) Proteins were separated by SDS-PAGE and transferred to a PVDF (Immobilon-P,
Merck Millipore, United States) membrane. The TMV cp antibody (Youlong,
Shanghai, China), anti-rabbit secondary antibody (CWBIO, Beijing,
China), and β-actin (CWBIO, Beijing, China) antibody were used
for this assay. Ubiquitination levels were then detected using antiubiquitin
as a primary antibody (PTM Biolabs, Hangzhou, China) and anti-rabbit
secondary antibody conjugated to HRP (CWBIO, Beijing, China). The
results of the western blot were analyzed by Image J (Version 1.52a).
the TMV coat protein (cp) and tobacco ubiquitination levels, proteins
were extracted from tobacco seedlings that had been treated with PBS
and TMV solution. Samples were collected at different time points.
We performed the western blot analysis using a commercially available
antibody against ubiquitination as previously described.32 (link),63 (link) Proteins were separated by SDS-PAGE and transferred to a PVDF (Immobilon-P,
Merck Millipore, United States) membrane. The TMV cp antibody (Youlong,
Shanghai, China), anti-rabbit secondary antibody (CWBIO, Beijing,
China), and β-actin (CWBIO, Beijing, China) antibody were used
for this assay. Ubiquitination levels were then detected using antiubiquitin
as a primary antibody (PTM Biolabs, Hangzhou, China) and anti-rabbit
secondary antibody conjugated to HRP (CWBIO, Beijing, China). The
results of the western blot were analyzed by Image J (Version 1.52a).