Fig. 5

mRNA and protein levels of CsPK at various life stages of C. sinensis. a Real- time PCR. The β-actin mRNA of C. sinensis was used as an internal control. Semiquantitative analysis was conducted using the 2^-ΔΔCt method. Significant differences in the mRNA levels of CsPK in egg, adult, excysted metacercaria, and metacercaria were observed (P < 0.01). The mRNA level of CsPK in egg was higher than that in adult (56.79-fold, t ₍₂₎ = 17.392, P = 0.003), metacercaria (9.72-fold, t _(2.02) = 15.844, P = 0.004) and excysted metacercaria (5.97-fold, t ₍₄₎ = 14.477, P < 0.001). b Western blotting. Total protein (40 μg) in extracts obtained at each life stage was probed with mouse anti-rCsPK sera, revealing specific immunoreactive protein bands at approximately 53.1 kDa. No corresponding band was detected with preimmune mouse serum (data not shown). c Relative protein levels were analysed using Tanon Gis software. The protein level of CsPK was maximal in eggs, followed by excysted metacercaria, metacercaria, and adults. The protein levels were consistent with the mRNA levels. (*P < 0.05; **P < 0.01; egg vs adult: t ₍₄₎ = 12.950, P < 0.001; excysted metacercaria vs adult: t ₍₄₎ = 16.542, P < 0.001; metacercaria vs adult: t ₍₄₎ = 13.951, P < 0.001; excysted metacercaria vs metacercaria: t ₍₄₎ = -3.680, P = 0.021)