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. 2015 Mar 10;10(3):e0119439. doi: 10.1371/journal.pone.0119439

Fig 7. Cloning and bioactivity determination of the GL18769 protein.

Fig 7

(A) The PCR product of GL18769 coding sequence. Total RNA was isolated from G. lucidum fruiting bodies (60 days) and amplified by reverse transcription-PCR. The GL18769 coding sequence (339bp) was amplified by PCR. (B) Expression and purification of GL18769 protein. A strong protein band appeared at approximately 13 kDa after IPTG-induction for 4 h compared with non-induction. This protein was purified using a His Trap FF column as indicated in the SDS-PAGE. (C) Stimulatory effect of GL18769 on mouse spleen lymphocytes. Compared to the ConA (2 μg/ml) treatment, GL18769 could significantly enhance the proliferation of mouse spleen lymphocytes (MSLs) in a dose-dependent manner (2.5, 5 and 10 μg/ml). (D) Representative morphology images of the MSLs treated with different dose of GL18769 (2.5, 5 and 10 μg/ml) after 36 hours. The results are means ± S.E.M. (n = 3); * P<0.05, ** P<0.01, *** P<0.001.