It was shown that the transformation efficiency of the test group of U937 cells (expressing Mtb Hsp16.3) was 73% (Figure 1A), and that of the control group (expressing GFP) was 82% (Figure 1B). Furthermore, to validate the expression of Mtb Hsp16.3 Selleck MCC 950 protein in the cells, western blot analysis was performed using anti-Mtb Hsp16.3 and the results demonstrated that Mtb Hsp16.3 was strongly expressed in the test group of U937 cells (Figure 1C). Figure 1 The integrase-deficient lentivirus vector (IDLV) transfected U937 cells with high efficiency and
the cells expressed Mtb Hsp16.3. An IDLV delivered the transgene into U937 histone deacetylase activity macrophages for instantaneous expression. The fluorescence microscopy and flow cytometry were used at 64 h after infection to detect GFP and analyse the transduction efficiency. A, the transduction efficiency of the test group of U937 cells (expressing Mtb Hsp16.3 and GFP) was 73%. B, the transduction efficiency
of the control group (expressing GFP only) was 82%. C, western blot analysis with antibodies against Mtb Hsp16.3; β-actin was used as a loading control. Expression profiles of miRNAs in U937 cells from the test group and the control group To determine the miRNA profiles for the two groups, the Exiqon miRCURY™ LNA Array was employed to perform the 2043 miRNAs assay (1898 human C188-9 research buy and 145 human viral miRNAs represented in the Sanger miRBase v18.0). After normalization and unsupervised filtering (see Methods), the obtained average values for each miRNA spot were used for statistical analysis. Comparing the data from the two groups (test/control) and using fold change filtering (upregulated more than 2-fold and downregulated less than 0.5-fold ), total of 149 differentially expressed miRNAs was identified, of which 60 were upregulated (Table 1) and 89 were downregulated (Table 2). hsa-miR-2355-3p 2.00 0.00162 2 hsa-miR-133b 4.30 0.00992 6 hsa-miR-451a 2.20 0.01085 17 hsa-miR-4664-3p 4.31 0.00022 8 hsa-miR-130b-3p 2.30 0.04627 22 hsa-miR-4431 4.35 0.00368 2 hsa-miR-486-5p Urocanase 2.32 0.00208 8 hsa-miR-4804-3p 4.36 0.00023 5 hsa-miR-361-5p 2.33 0.04722 X hsa-miR-18b-3p 4.62 0.00191 X hsa-miR-3156-3p 2.50 0.00729 10 hsa-miR-675-3p 4.68 0.00028 11 hsa-miR-4728-3p 2.67 0.00029 17 hsa-miR-550b-3p 4.72 0.01382 7 hsa-miR-3191-5p 2.67 0.00020 19 hsa-miR-551a 4.75 0.00063 1 hsa-miR-296-5p 2.71 0.04951 20 hsa-miR-4685-3p 5.04 0.00090 10 hsa-miR-150-5p 2.85 0.00927 19 hsa-miR-23c 5.11 0.00081 X hsa-miR-4540 2.86 0.01280 9 hsa-miR-5002-3p 5.14 0.00035 3 hsa-miR-4268 2.97 0.00969 2 hsa-miR-5689 5.33 0.00054 6 hsa-miR-1236 3.08 0.04877 6 hsa-miR-935 5.43 0.00187 19 hsa-miR-221-5p 3.16 0.03132 X hsa-miR-374b-3p 5.79 5.