Fig. 4

clr-2 overexpression leads to increased cellulase when complemented with increased Ca²⁺ in the growth medium. RT‒PCR showing transcript levels of a clr-2 and b cellulase genes when expressed under native, gpd (constitutive), and cbh1 (inducible) promoters. c Total cellulase activity of secretomes obtained when clr-2-overexpressing strains were grown in RCM medium. d Total cellulase activity of secretomes obtained when P_clr2Clr-2/∆Mig1 was grown in RCM medium with varying levels of CaCO₃. Secretome profile on 12% SDS‒PAGE e and total cellulase activity f by FPU assay of clr-2 mutants grown in RCM medium with 2.5 g/L and 5.0 g/L CaCO₃. All experiments were repeated three times; statistical significance was determined by a one-tailed, unequal variance t-test