TABLE 2.
Expression of autophagy genes in BM-derived infected macrophages from transgenic and WT micea
| Gene | Fold change (mean ± SEM) |
||
|---|---|---|---|
| M1-γ34.5 | M2-γ34.5 | WT | |
| Akt1e | P > 0.05 | P > 0.05 | P > 0.05 |
| Akt2 | P > 0.05 | P > 0.05 | 2.22 ± 0.54*f |
| Akt3e | P > 0.05 | P > 0.05 | P > 0.05 |
| Atg10e | P > 0.05 | P > 0.05 | P > 0.05 |
| Atg101e | P > 0.05 | P > 0.05 | P > 0.05 |
| Atg12d | 1.26 ± 0.01 | 1.48 ± 0.03 | 1.51 ± 0.04 |
| Atg13d | 1.70 ± 0.10 | 1.76 ± 0.03 | 2.11 ± 0.14 |
| Atg3d | 1.64 ± 0.06 | 1.57 ± 0.02 | 1.37 ± 0.05 |
| Atg4ad | 1.18 ± 0.07 | 1.31 ± 0.02 | 1.2 ± 0.05 |
| Atg4bd | 1.58 ± 0.11 | 1.45 ± 0.05 | 1.41 ± 0.09 |
| Atg5e | P > 0.05 | P > 0.05 | P > 0.05 |
| Atg7e | P > 0.05 | P > 0.05 | P > 0.05 |
| Atg9a | 1.88 ± 0.17* | 1.59 ± 0.06* | P > 0.05 |
| Badd | 1.50 ± 0.11 | 1.64 ± 0.09 | 1.70 ± 0.14 |
| BclIIe | P > 0.05 | P > 0.05 | P > 0.05 |
| Bcl2l1d | 1.81 ± 0.07 | 1.63 ± 0.08 | 1.57 ± 0.03 |
| Becn1d | 1.23 ± 0.03 | 1.55 ± 0.05 | 1.40 ± 0.01 |
| Bnip3b | −2.32 ± 0.09 | −1.56 ± 0.10 | −4.72 ± 1.9 |
| Cflarc | 7.22 ± 0.37 | 5.31 ± 0.38 | 4.06 ± 0.26 |
| Ctsdd | −1.89 ± 0.01 | −1.68 ± 0.04 | −1.66 ± 0.03 |
| Ctsle | P > 0.05 | P > 0.05 | 2.06 ± 0.38* |
| Ddit4e | P > 0.05 | P > 0.05 | P > 0.05 |
| Eif2ak3d | −1.32 ± 0.15 | −1.50 ± 0.09 | −1.50 ± 0.09 |
| Ern1d | 1.56 ± 0.05 | 1.66 ± 0.11 | 1.55 ± 0.12 |
| Hif1a | 2.72 ± 0.44* | P > 0.05 | P > 0.05 |
| Hmgb1 | −1.52 ± 0.07 | −2.66 ± 0.6 | −2.95 ± 0.67 |
| Hrasd | 1.34 ± 0.07 | 1.36 ± 0.04 | 1.38 ± 0.05 |
| Krasd | 1.85 ± 0.23 | 1.92 ± 0.19 | 2.20 ± 0.14 |
| Lamp1d | 1.07 ± 0.02 | 1.51 ± 0.03 | 1.28 ± 0.02 |
| Lamp2d | 1.39 ± 0.04 | 1.75 ± 0.03 | 1.69 ± 0.06 |
| Map2k1d | 1.91 ± 0.1 | 1.85 ± 0.09 | 1.63 ± 0.09 |
| Map2k2d | 2.01 ± 0.16 | 1.60 ± 0.17 | 1.58 ± 0.14 |
| Map3k7e | P > 0.05 | P > 0.05 | P > 0.05 |
| Mapk1e | P > 0.05 | P > 0.05 | P > 0.05 |
| Mapk3d | −1.31 ± 0.05 | −1.38 ± 0.14 | −1.26 ± 0.06 |
| Mapk8e | P > 0.05 | P > 0.05 | P > 0.05 |
| Mapk9d | 1.72 ± 0.03 | 1.89 ± 0.06 | 1.53 ± 0.04 |
| Mtmr14 | 1.65 ± 0.09* | P > 0.05 | P > 0.05 |
| Mtmr3 | 1.37 ± 0.08* | P > 0.05 | P > 0.05 |
| mTOR | 1.39 ± 0.18* | P > 0.05 | P > 0.05 |
| Nrase | P > 0.05 | P > 0.05 | P > 0.05 |
| Nrbf2 | P > 0.05 | 1.29 ± 0.12* | P > 0.05 |
| Pik3c3e | P > 0.05 | P > 0.05 | P > 0.05 |
| Pik3cae | P > 0.05 | P > 0.05 | P > 0.05 |
| Pik3cbe | P > 0.05 | P > 0.05 | P > 0.05 |
| Pik3cd | P > 0.05 | P > 0.05 | 4.00 ± 1.13* |
| Pik3r2e | P > 0.05 | P > 0.05 | P > 0.05 |
| Pik3r3e | P > 0.05 | P > 0.05 | P > 0.05 |
| Pik3r4e | P > 0.05 | P > 0.05 | P > 0.05 |
| Prkcd | 1.44 ± 0.05* | 1.37 ± 0.09* | P > 0.05 |
| Prkcqe | P > 0.05 | P > 0.05 | P > 0.05 |
| Ptene | P > 0.05 | P > 0.05 | P > 0.05 |
| Raf1d | 1.84 ± 0.08 | 1.76 ± 0.14 | 1.59 ± 0.13 |
| Rb1cc1 | P > 0.05 | 1.20 ± 0.05* | P > 0.05 |
| Rps6kb1d | 1.30 ± 0.11 | 1.43 ± 0.03 | 1.38 ± 0.04 |
| Rras2 | P > 0.05 | P > 0.05 | −2.63 ± 0.51* |
| Stk11 | 1.25 ± 0.08* | P > 0.05 | P > 0.05 |
| Traf6d | 2.17 ± 0.06 | 1.94 ± 0.27 | 1.87 ± 0.21 |
| Tsc2e | P > 0.05 | P > 0.05 | P > 0.05 |
| Ulk1e | P > 0.05 | P > 0.05 | P > 0.05 |
| Ulk2 | −1.52 ± 0.11* | P > 0.05 | P > 0.05 |
| Wipi1 | −1.68 ± 0.13* | −2.26 ± 0.83* | P > 0.05 |
BM-derived macrophages from M1-γ34.5 and M2-γ34.5 transgenic mice and WT mice were infected with 10 PFU/cell of WT McKrae virus for 1 h at 37°C, washed with PBS, and incubated for an additional 24 h in fresh medium. Expression of 62 autophagy genes in infected cells was analyzed by NanoString, and the level of each gene was normalized to its level in mock-infected BM-derived macrophages. Results are shown as fold increase or decrease relative to mock-infected gene expression. Results indicate mean ± SEM (n = 3).
For Bnip3 gene expression, there were no differences between the WT and M1-γ34.5 groups (P > 0.05), while differences between WT and M2-γ34.5 groups and among WT, M1-γ34.5, and M2-γ34.5 were statistically significant (P < 0.05).
For Cflar gene expression, the three groups of infected macrophages were statistically significantly different from each other (P < 0.05).
The three groups of infected macrophages were not significantly different from each other (P > 0.05), but they were significantly different from their mock-infected counterparts (P < 0.05).
M1-γ34.5, M2-γ34.5, and WT macrophages were not significantly different from each other or their mock-infected counterparts for AKT1, AKT3, AKT10, Atg101, Atg5, Atg7, BclII, Ddit4, Map3k7, Mapk1, Pik3c3, Pik3ca, Pik3cb, Pik3r2, Pik3r3, Pik3r4, Prkcq, Pten, Tsc2, and Ulk1 genes (P > 0.05).
Each gene indicated by an asterisk among the three groups of macrophages was similar to one another and significantly different from their counterparts without the asterisk.