Novel insights into immune-related genes associated with type 2 diabetes mellitus-related cognitive impairment

Figure 10  Validation of the expression of hub genes in vivo and in vitro. A: Hub genes mRNA expression in db/db and db/m mice by reverse transcription quantitative real-time PCR (RT-qPCR). In comparison to db/m mice, H2-T24 and Rac3 showed a reduction in expression in db/db mice, whereas Tfrc demonstrated an elevation in expression in db/db mice; B: Palmitic acid (PA) was used as a type 2 diabetes mellitus model in vitro. Hub genes mRNA expression in PA and negative control (NC) groups by RT-qPCR. H2-T24 and Rac3 showed a reduction in expression in the PA group, whereas Tfrc demonstrated an elevation in expression in the PA group; C: Protein levels of RAC3 and TFRC in db/db and db/m mice by western blotting. Each lane represents an independent mouse hippocampus; D: Analysis of relative protein levels in db/db and db/m mice. In comparison to db/m mice, Rac3 showed a reduction in expression in db/db mice, whereas Tfrc demonstrated an elevation in expression in db/db mice; E: Protein levels of RAC3 and TFRC in PA and NC groups by western blotting. Each lane represents an independent experiment of cell lysate; F: Analysis of relative protein levels in PA and NC group. Rac3 showed a reduction in expression in the PA group, whereas Tfrc demonstrated an elevation in expression in the PA group; G: Protein expression of RAC3 and TFRC in db/db and db/m mice by immunohistochemistry; H: Analysis of average OD in db/db and db/m mice. In comparison to db/m mice, Rac3 showed a reduction in expression in db/db mice, whereas Tfrc demonstrated an elevation in expression in db/db mice. Data are expressed as the mean ± SEM (n = 3). ^aP < 0.05; ^bP < 0.01; ^cP < 0.001. NC: Negative control; PA: Palmitic acid.