*2.2. Antibodies*

The following primary antibodies were used: mouse GAPDH (ab9484), rabbit β-tubulin (ab6046) from Abcam (Cambridge, UK); rabbit phospho-Akt Ser473 (D9E XP), phospho-p38 MAPK (T180/Y182), phospho-STAT1 (Tyr701) (58D6), phospho-TAK1 (T184/187) (90C7), phospho-JNK (81E11) (T183/Y185), phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204), IκB α (44D4), IRAK1 (D51G7), MyD88 (D80F5), Histone H3 (3H1), and phospho-NF-κ<sup>B</sup> p65 (Ser536) (93H1) from Cell Signaling Technology (Danvers, MA, USA); rabbit PCNA Abs were from Santa Cruz Biotech (Santa Cruz, CA, USA); sheep IRF5 and IRAK4 were from MRC-PPU Reagents (University of Dundee, Dundee, UK); goa<sup>t</sup> TIRAP polyclonal Abs were from Invitrogen (#PA5-18439, Waltham, MA, USA), and mouse STAT1 antibodies were from BD Biosciences (#610185, Wokingham, UK). Secondary antibodies (HRP-linked) were from DAKO Denmark A/S (Glostrup, Denmark).

### *2.3. siRNA Treatment*

Oligos used for silencing were AllStars Negative Control siRNA (SI03650318) and FlexiTube Hs\_TIRAP\_10 siRNA (SI03075135) (QIAGEN, Germantown, AR, USA). PBMCs were seeded in 24-well plates (NUNC, ThermoFisher Scientific, Waltham, MA, USA), 1.5 × 10<sup>6</sup> cells per well, and differentiated to MDMs as described above. On day 7 and day 9, cells were transfected by silencing and control oligo (20 nM final concentration) using Lipofectamine 3000 (Invitrogen, ThermoFisher Scientific, Waltham, MA, USA), as suggested by the manufacturer. Cells were stimulated by LPS, CL075 or FSL-1, or used for bacterial infections in 48 h after second transfection.
