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Figure 6 | BMC Biochemistry

Figure 6

From: Effect of polyamines and synthetic polyamine-analogues on the expression of antizyme (AtoC) and its regulatory genes

Figure 6

a Effect of polyamines or polyamine analogues on endogenous DnaK expression. Western blot of total cell extracts of E. coli K12 cells BW25113 (atoSC+) grown to an OD600 0.8–0.9 (15 μl), was performed to detect the expression of the heat shock protein DnaK. Lane 1: BW25113 cells with no inducer. Lanes 2–4: BW25113 cells treated with PUT, SPD, or SPM, respectively. Lanes 5–9: BW25113 cells treated with 1 mM polyamine analogue 2, 10, 12, 13, 14, respectively. 6 b. Effect of polyamines or polyamine analogues on endogenous AtoC/Az expression. The E. coli K12 cells BW25113(atoSC+) were cultured in the presence of 1 mM of the indicated polyamine analogue or each polyamine, respectively. Total cell extracts of each culture grown to an OD600 0.8–0.9 (20 μl) were analyzed. The immunoblot analysis was performed using the purified polyclonal rabbit antibody of AtoC/Az. BW28878 cells, not expressing the atoC/Az protein, were used as negative control. The high copy plasmid pUC-Az [21], overexpressing the AtoC/Az protein, was used as a positive control. Lane 1: BW28878 cells, Lane 2: BW25113 cells with no inducer Lanes 3–5: BW25113 cells treated with PUT, SPD or SPM, respectively. Lanes 6–9: BW25113 cells treated with analogues 10, 12, 14, 13. Lane 10: BW25113 cells carrying the pUC-Az plasmid.

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