Fig. 6

DNAs in T-MPs stimulate monocytes to upregulate IRF4 expression. a BALB/c mouse monocytes were co-incubated with PKH26-stained H22-MPs, cytochalasin D or PKH26-stained H22-MPs mixed with cytochalasin D. After 2, 4, 6 or 12 h, PKH26 fluorescence intensity of monocytes was measured via flow cytometry, and the IRF4 expression was analyzed at the levels of both gene and protein. b Monocytes pretreated with or without 2-DG for 30 min were co-incubated with H22-MPs for 24 h or 48 h. IRF4 expression was measured at the levels of both gene and protein. c Monocytes were co-cultured with PKH26 or PKH67-labeled H22-MPs for 14 h. Then the monocytes were analyzed with ER, Golgi, mitochondria, endosome or lysosome fluorescent trackers using a two-photon confocal microscope. d Monocytes were treated with DNAs or RNAs extracted from H22-MPs. After 24 or 48 h, the IRF4 expression was analyzed at the levels of both gene and protein. e Monocytes were co-cultured with H22-MP-derived DNAs or H22-MP-derived DNAs pretreated with DNase. 24 or 48 h later, IRF4 expression was analyzed at the levels of both gene and protein. f Monocytes transfected with TLR9 siRNA were co-cultured with H22-MPs. After 24 or 48 h, IRF4 expression was analyzed at the levels of both gene and protein. Mean ± s.e.m. is represented in the data and two-tailed unpaired Student's t test was used to statistically analyze the P values. **P < 0.01; ***P < 0.001; NS, not significant