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. 2019 Feb 4;116(8):3042–3051. doi: 10.1073/pnas.1811589116

Fig. 8.

Fig. 8.

The influence of SraL in Rho protein expression and activity. (A) Detection of Rho protein under denaturing conditions was performed by Western blot from 5 μg of total protein extracts obtained in middle exponential phase, stationary phase, and upon anaerobic shock. Dashed lines indicate noncontiguous lanes. (B) Native Western blot to evaluate Rho protein expression in middle exponential phase was done from 40 μg of total protein cellular extracts. For both denaturing and native conditions Rho expression was determined in strains expressing SraL sRNA at different levels using anti-Rho antibody (MyBioSource). (C) β-Galactosidase activity was accessed in a strain carrying a fusion of LacZ reporter under the control of pgaABDC operon promoter and rut site, using o-nitrophenyl-β-d-galactopyranoside (Sigma) as substrate. Rho protein activity in pgaABDC promoter was evaluated in middle exponential phase, stationary phase, and upon anaerobic shock. Results are expressed in Miller units and are representative of at least three biological replicates. Statistical analysis was carried out using GraphPad Prism 6 software. ****P < 0.001 (Paired t test).