Figure 1From: Establishment of a mouse model to express bovine CD14 short hairpin RNA Screening of bovine CD14 shRNA and construction of its eukaryotic expression vector. A. Effect of designed bovine CD14 shRNAs was detected by qRT-PCR analysis. The lentiviral particles expressing CD14 shRNAs were used to infect HEK 293 cells expressing bovine CD14, non-infected cell line as a blank control, the scrambled shRNA as negative control. The values for columns with different letters represent statistically significant differences, p < 0.01. B. The inhibition effect of CD14 shRNA-674 fragment was confirmed by western blot analysis. HEK 293 cells stably expressing bovine CD14 were infected by shRNA-674 lentivirus at two different MOIs (lane1, 2), the negative control was HEK 293 cells stably expressing bovine CD14 (lane 3). C. Identification of the pSilencer™4.1-CD14-IRES recombinant plasmid. M: 1 kb Marker; Lane1: pSilencer™4.1-CD14shRNA-IRES plasmid: Lane2: pSilencer™4.1-CD14shRNA-IRES plasmid digested by SspI enzyme; Lane3: pSilencer™4.1-CD14shRNA-IRES plasmid digested by HpaI and BamHI enzyme. D. Map of pSilencer™4.1-CD14 shRNA-IRES vector. E. Confirmation of F1 generation transgenic mice by Southern blot analysis. Three F1 mice were selected, among them, two (offspring of 22#, 32#) were transgenic while the third was not (offspring of 11#) by RT-PCR analysis. The positive control was pSilencer™4.1-CD14 shRNA-674-IRES plasmid. M: Marker; Lane 1: positive control; Lane 2: offspring of 22# mouse; Lane 3: offspring of 11# mouse; Lane 4: offspring of 32# mouse.Back to article page