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. 2013 Feb 20;8(2):e56888. doi: 10.1371/journal.pone.0056888

Figure 2. Detection of MD-2 by Western blot in HT-29 cells infected with C. concisus strains.

Figure 2

HT-29 cells were lysed following incubation with C. concisus strains for 24 hours. Expression of MD-2 in HT-29 cells was detected by Western blot. Two bands, the Glycosylated MD-2 and non-glycosylated MD-2, were revealed on Western blot. Giving the close distance of Glycosylated MD-2 and non-glycosylated MD-2 protein bands which made it difficult to analyze the bands separately, these two protein bands were analyzed together. The level of MD-2 was expressed as the fold change of the normalized band intensity of a sample relative to the normalized band intensity of the negative control (HT-29 cells without C. concisus infection), after normalization to the intensity of the internal control α-Tubulin of the same sample. A: Representative Western blot of MD-2 (23–25 kD) and α-Tubulin (55 kD). B: Level of total MD-2 induced by different C. concisus strains; data were the average of triplicate experiments ± standard error. N: negative control. H2O1-H6O1: C. concisus strains isolated from healthy controls. The remaining nine C. concisus strains were from patients with IBD. The average level of MD-2 induced by C. concisus strains from patients with IBD was not significantly higher than that induced by C. concisus strains from healthy controls (P>0.05).