Table 1

The siRNA sequence of GBP2b and negative control"

Table 2

The real-time PCR primers used in this study"

Table 3

Antibody information"

Fig. 1

Changes in mRNA and protein levels of GBP2b after M. bovis infected RAW264.7 cells Detection of GBP2b mRNA and protein expression after infection of RAW264.7 cells with M. bovis at MOI of 0, 2, 10, and 50 for 24 h, or infected with M. bovis at MOI of 10 for 0, 6, 12, and 24 h, respectively. A, B. Results of qRT-PCR analysis of GBP2b in RAW264.7 cells; C, D. Results of Western blot analysis of protein expression of GBP2b in RAW264.7 cells. Values are "${\bar x}$±s" for at least three independent experiments performed in triplicate. ***. P < 0.001. MOI. The multiplicity of infection; HPI. Hours post-infection"

Fig. 2

GBP2b interference effect The mRNA and protein expression levels of GBP2b were detected by qRT-PCR(A) and Western blot(B) after infection of RAW264.7 cells with 10 MOI M. bovis. Values are "${\bar x}$±s" for at least three independent experiments performed in triplicate. *P < 0.05, *** P < 0.001, ns. No significance"

Fig. 3

Detection of M1 and M2 macrophage markers following GBP2b downregulation during M. bovis infection A, B. The mRNA expression levels of M1 marker genes (IL-1β, iNOS, and TNF-α) and M2 marker genes (IL-10, Arg1, and Mrc1) in RAW264.7 cells interfering with GBP2b were detected by qRT-PCR; C. Flow cytometry detection of CD86 or CD206 positive cells in RAW264.7 cells interfered with GBP2b.Values are "${\bar x}$±s" for at least three independent experiments performed in triplicate. *.P < 0.05, **.P < 0.01, ***. P < 0.001, ns. No significance"

Table 4

Prediction results of binding sites of the GBP2b and NF-κB promoters"

Fig. 4

Double luciferase activity detection of NF-κB transcription"

Fig. 5

Expression of TLR-related proteins after M. bovis infection with RAW264.7 cells that interfered with GBP2b."

Fig. 6

Immunofluorescence detection of NF-κB p65 after M. bovis infection interfered with GBP2b"