Additional file 2 of PotD contributes to Streptococcus suis-induced blood–brain barrier disruption by regulating arcA transcription

Additional file 2. Results of PCR validation of ΔpotD and CΔpotD and purification of recombinant PotD protein and ADI protein. A, B PCR verification of ΔpotD mutant (A) and complementary mutant strain CΔpotD (B). C Purification of His-tagged PotD protein. Lane M: Protein marker (180 kDa); Lane 1: Unpurified cell lysate; Lane 2: Proteins removed with 10 mM imidazole buffer; Lane 3: Proteins removed with 20 mM imidazole buffer; Lane 4: His-PotD protein eluted with 250 mM imidazole buffer. D Purification of GST-tagged ADI protein. Lane M: Protein marker (250 kDa); Lanes 1–8: GST-fused protein eluted with 10 mM reduced glutathione in elution buffer. The target proteins are denoted by red boxes. E SDS–PAGE analysis of total soluble protein extract of SC19 and purified His-PotD. Lane M: Protein marker (180 kDa); Lane 1: Total soluble protein extract of SC19; Lane 2: Purified His-PotD; Lane 3: Mixture of total soluble protein extract of SC19 and His-PotD. The excised gel band (Lane 3) was subjected to mass spectrometry analysis.