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Figure S1: Effects of ATF6 siRNA. (A) ATF6 siRNA selectively inhibited the ATF6 expression in ER-stressed macrophages. The ATF6- or NS-siRNA transfected BMMs were pretreated with Tm, followed by stimulations with LPS, as described in the Materials and Methods section. Cells were harvested at 4 h to measure gene expressions of ATF6, XBP1, CHOP and Grp78 by qRT-PCR. (B) ER stress did not alter IP-10 expressions. BMMs were pretreated with Tm or Tg, followed by LPS stimulations for 4 h, as described in the Materials and Methods section. IP-10 gene expressions were measured by qRT-PCR. (C) ATF6 siRNA did not regulate macrophage IP-10 gene expression. As described in (A), IP-10 expression in siRNA transfected macrophages under ER stress was measured by qRT-PCR. Results are representative of at least two separate experiments, n = 3 replicates/group/expt., t-test, *p < 0.05.

Figure S2: Targeted delivery of siRNA in vivo by mannose-conjugated polymers. AlexaFluor488-labeled siRNA (green) were mixed with mannose-conjugated polymers and injected via tail vein, as described in the Materials and Methods section. Spleen and livers were collected at 4 h postinjection. Tissue specimens were fixed with 10% neutral formaldehyde, embedded and frozen in O.C.T (Tissue-Tek® optimum cutting temperature compound, VWR). Liver sections (4 µm) were stained with anti-CD68 antibody (Santa Cruz Biotechnology, Inc., San Diego, CA) and Cy3-conjugated AffiniPure goat anti-mouse IgG antibody (red; Jackson Immunoresearch, West Grove, PA). DAPI (blue) was used for nuclear counterstaining. The biodistribution of green fluoresence positive cells in spleen were indicative of phagocytes (expressing mannose receptors) being transfected. In livers, these positive cells merged with red immunofluorescence-stained CD68 positive cells.

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