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. 2016 Dec 26;7:1931. doi: 10.3389/fpls.2016.01931

FIGURE 2.

FIGURE 2

Western blot analysis and GUS assay. Total microspore protein were analyzed using western blotting with specific Anti-His HRP-conjugated antibodies (1:1000). (a,b) Confocal microscopy imaging of trypsin-EDTA (0.25%) washed microspores, red fluorescence denotes Bcl-2△21 labeled Alexa Fluor® 647 inside the microspore but absent in exine. Bars: 20 μM. (c) Western blotting with specific Anti-His HRP-conjugated antibodies (1:1000). M, prestained benchmark protein marker (ThermoFisher Scientific). 1, total protein extracted from untreated microspores. 2–4, total protein extracted from Bcl-2△21 treated microspores. The arrow denotes the presence of specific Bcl-2△21 bands. GUS histochemical staining using 5-bromo-4-chloro-3-indolyl glucuronide (X-Gluc) as the substrate in microspores after transduction. (d) GUS protein without R10HL; (e) GUS protein conjugated with R10HL; (f) GUS assay without R10HL (control), GUS protein conjugated with R10HL (transduced). All other conditions were as mentioned in Figure 1.