Journal: The Plant Journal
Article Title: Recovery of RNA ‐dependent RNA polymerase 6 gene‐knockout phenotypes in Nicotiana benthamiana via in vivo generation of inverted repeat construct of the trans‐acting short interference RNA3 sequence
doi: 10.1111/tpj.70350
Figure Lengend Snippet: Transient expression of GFP and human FGF1 in WT, TAS3i, and rdr6 plants. GFP and human FGF1 were transiently expressed in WT, TAS3i, and rdr6 plants, and the infiltrated leaves were harvested at 5 dpi. (a) The relative expression level of GFP mRNA was estimated by real‐time RT‐PCR using NAD(P)H dehydrogenase (quinone) ( NQO ) as the housekeeping gene. (b) The amount of GFP protein was estimated using SDS‐PAGE followed by Coomassie blue staining. (c) Relative GFP fluorescence was measured by 20‐fold diluting the centrifuged supernatant of the crude extract (prepared in PBS at five times the volume of the leaves) from GFP‐expressing leaves. (d) The relative expression level of FGF1 mRNA was estimated by real‐time RT‐PCR using NQO as the housekeeping gene. (e) Relative band intensities of FGF1 were estimated from Western blot analysis. Data are shown as mean ± standard deviation ( n = 3). t ‐test; * P < 0.05; n.s., not significant.
Article Snippet: To detect human FGF1 expression, anti‐FGF‐1 (B‐3), human (mouse) (Santa Cruz Biotechnology, Dallas, TX, USA, #SC‐55520) was used as the primary antibody.
Techniques: Expressing, Quantitative RT-PCR, SDS Page, Staining, Fluorescence, Western Blot, Standard Deviation
