Supplementary Figure 1 Rluc8 β- lactamase araC Ori from pMB1 pBAD-RLuc bp P BAD ClyA β- lactamase araC Ori from pMB1 pBAD-ClyA 4883 bp P BAD A B Supplementary Figure 2

Supplementary Figure 3 Supplementary Figure 4

Supplementary Figure 5 P1 & C2P2 & C1C1 & C2Primer pairs :P2 & C3 1. (Δara operon::cat ΔppGpp S. typhimurium) PCR templates 1.Δara operon::cat ΔppGpp S. typhimurium 2.ΔppGpp S. typhimurium 12 Templates : A B 1 kb 4 kb 1 kb 1.5 kb 0.5 kb

Supplementary Figure 1: Plasmid maps of pBAD-RLuc8 (pRLuc8) (A) and pBAD-ClyA (pClyA) (B). Supplementary Figure 2: Semi-quantitative densitometry of the secretion efficiencyof ClyA in Westernblot analysis (Figure 4). An arbitrary unit of band from filtrate was divided by that of band from bacterial pellet and it was calculated as a percentage of secreted protein (%) Supplementary Figure 3: Changes in the body weight of CT26 tumor bearing mice after intravenous injection of ΔppGpp S. typhimurium (SL) or ara operon deleted S. typhimurium (SL ΔARA ), both of which are expressing ClyA. CT26 tumor bearing mice were injected I.V. with the bacteria (3.0 × 10 7 CFU of SL-pClyA or SL ΔARA -pClyA, n=5 each group). The body weight changes were compared with the untreated mice. The body weight of CT26 tumor bearing mice was not significantly different between untreated and treated mice. The experiment was terminated when the tumor volume exceeded 1,500 mm 3. Supplementary Figure 4: Number of viable S. typhimurium (SL) and ara operon deleted S. typhimurium (SL ΔARA ) ex vivo at day 5, 8, 10, and 15 after bacterial injection of CT26-bearing BALB/c mice. Number of CFU per gram of tissue was counted. n = 3 per group. NS, no statistical difference. Supplementary Figure 5: PCR verification of deleted ara operon and replacement of chloramphenicol acetyltransferase gene. (A) Δara operon::cat ΔppGpp S. typhimurium was used as a PCR template with each pair of primers (P1 and C2 ; P2 and C1 ; C1 and C2). (B) Both strains (lane 1: Δara operon::cat ΔppGpp S. typhimurium, lane 2 : ΔppGpp S. typhimurium) were used as a PCR template with primer pairs (P2 and C3).