Fig 2: Characterization of AA mouse livers homogenates. (A) Total 2 µg of homogenates from AA-mL (lane i) and AA-mLdec (lane ii) were separated in SDS 15% PAGE under reducing conditions. Apparent molecular weights for the marker are given in kDa. 1 and 2 indicate bands subjected to mass mapping analysis. (B) Tryptic peptides obtained by digestion of SDS-PAGE bands were analyzed by MALDI-MS. MH+ monoisotopic values (Da) are reported for each peptide. (C) Western blot analysis of AA-mL (i) and AA-mLdec (ii) and standard serum mouse SAA1 (11.6 kDa) from the same strain of double transgenic mice [12] as AA-mL and AA-mLdec.

Fig 3: Gene contents of SAA1, TLRs, E3 Ligases, inflammatory cytokines and NF?B in soleus, plantaris, and EDL muscles. (a) Gene contents in soleus muscle. (b) Gene contents in plantaris muscle. (c) Gene contents in EDL muscle. Lettering denotes statistical significance with alpha set at 0.05. For all RT-PCR analysis an n of 8 samples per group was used

Fig 4: SAA1-4 expression in mouse small intestinal organoids upon stimulation with several cytokines.Normalized SAA expression levels in small intestinal organoids upon stimulation with (A) IL-6 (10 ng/mL), (B) TNF-a (10 ng/mL), (C) IL-1ß (100 ng/mL), (D) IL-23 (50 ng/mL), (E) IL-22 (50 ng/mL), or (F) IL-10 (50 ng/mL) for 24 h. Only IL-6, TNF-a, and IL-22 promoted a significant increase in SAA1 expression level when compared with untreated controls. Although not statistically significant, the expression level of SAA1 tended to increase upon IL-1ß stimulation and was suppressed by stimulation with IL-10. It was difficult to evaluate SAA2 and SAA4 expression because their expression levels were not detectable (A-F). The expression level of SAA3 was significantly increased by IL-22 and IL-23 (D, E). However, unexpectedly, SAA3 was downregulated by IL-1ß (C). SAA expression level was normalized to that of ACTB and is represented as the mean ± standard deviation (n = 5). **P < 0.01 versus untreated controls. N.S., not significant; N.D., not detected.

Fig 5: Induction of proinflammatory factors in placenta and fetal membranes by intraperitoneal administration of LPS or SAA1 in the mouse. (A,B) Induction of SAA1/2 expression in mouse placenta (A) and fetal membranes (B) by intraperitoneal administration of LPS (5 mg/kg BW, 6 h) at gestational day 16.5 (n = 3). (C–E) Induction of IL-1ß (C), TNF-a (D) and COX-2 (E) in the mouse placenta by intraperitoneal administration of LPS (5 mg/kg BW, 6 h) and SAA1 (8 µg/kg BW, 6 h) at gestational day 16.5 (n = 4). Data are mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001.