Skip to main content
. 2020 Aug 19;6(3):141. doi: 10.3390/jof6030141

Figure 2.

Figure 2

Expression of the PAFC encoding gene and protein. Total RNA was extracted from (A) mycelium grown in submersed and (B) synchronized surface cultures up to 96 h. Ten μg of total RNA was loaded per lane on a 1.2% denaturing agarose gel, blotted onto a nylon membrane and hybridized with a pafC specific digoxigenin-labelled PCR probe (upper panels). Ethidium bromide-stained 26S and 18S rRNA provide loading controls (lower panels). (C) For Western blot analysis, 20 μL of the cell free supernatant from submersed cultures was loaded per lane, size fractionated on an 18% (w/v) SDS-polyacrylamide gel and transferred onto a nitrocellulose membrane. A polyclonal antibody generated against PeAfpC was used to probe for PAFC. Recombinant PAFC (0.5 μg) was used as a control for antibody binding. The black arrow indicates PAFC (6.6 kDa). LRM, low range rainbow marker (GE Healthcare Life Sciences, Little Chalfont, UK). The molecular weight marker bands of 3.5 kDa and 8.5 kDa are indicated.