Phospho-IRF3 (Ser386) Antibody - #AF3438

Figure 6. Ca2+-dependent MLC activation enhances filopodia formation via p-cortactin rearrangement. (A) Representative immunofluorescence (IF) images of MKN-45 and DLD-1 cells stained for mitochondria and F-actin (red: mitochondria, green: F-actin). Scale bar: 10 μm. (B) Representative MYO10 IF staining in MKN-45 and DLD-1 cells treated with CCCP (10 μM) (blue: nuclei, red: MYO10, green: F-actin). Scale bar: 20 μm. (C) Quantification of MYO10-positive filopodia in MKN-45 and DLD-1 cells treated with CCCP (10 μM). Data are expressed as means ± SD, **P < 0.01, ***P < 0.001 (vs. DMSO group). (D) Representative IF staining of MKN-45 and DLD-1 cells (blue: nuclei, red: dil, green: MYO10, violet: p-cortactin). Scale bar: 20 μm (left). Zoomed-in images and fluorescence co-localization of p-cortactin and MYO10 at filopodia tips in MKN-45 and DLD-1 cells (MKN-45: Pearson > 0.9; DLD-1: Pearson > 0.9). Scale bar: 2 μm (right). (E) Quantification of p-cortactin-positive filopodia tips per cell in MKN-45 and DLD-1 cells treated with CaCl2 (5 mM) or BAPTA-AM (10 μM). Data are expressed as means ± SD, *P < 0.05, **P < 0.01 (vs. DMSO group). (F) Quantification of p-cortactin-positive filopodia tips per cell in MKN-45 and DLD-1 cells treated with calyculin A (50 nM) or P18 (8 μM). Data are expressed as means ± SD, *P < 0.05, **P < 0.01, ***P < 0.001 (vs. DMSO group). (G) Representative MYO10 IF staining in MKN-45 and DLD-1 cells treated with PP2 (10 μM) (blue: nuclei, red: MYO10, green: F-actin). Scale bar: 20 μm. (H) Quantification of MYO10-positive filopodia in MKN-45 and DLD-1 cells treated with PP2 (10 μM). Data are expressed as means ± SD, *P < 0.05, **P < 0.01 (vs. DMSO group).

Fig. 6: TLR3 signal activation induces HAO1 expression in alveolar epithelial cells. A Effect of 67NR and 4T1 exosomes treatment on HAO1 expression in alveolar epithelial cells by Western blot analysis. B Effect of transfection of RNA isolated from 67NR and 4T1 exosomes on HAO1 expression in alveolar epithelial cells by Western blot analysis. C Effect of poly (I:C) treatment on HAO1 expression in alveolar epithelial cells by Western blot analysis. D Effect of poly (I:C) inhalation on HAO1 expression in mice lung by Western blot analysis. E Detection of oxalate production in alveolar epithelial cells treated with poly(I:C) or poly(I:C) + CCPST. Means ± s.e.m are provided (n = 3). F Detection of TLR3 expression in alveolar epithelial cells infected with sg-control or sg-Tlr3 lentivirus. G Detection of HAO1 expression in alveolar epithelial cells treated with 4T1 exosomes, 4T1 exosomes + CU CPT 4a or 4T1 exosomes + sg-Tlr3 lentivirus by Western blot analysis. H Detection of oxalate production in alveolar epithelial cells treated with 4T1 exosomes, 4T1 exosomes + CU CPT 4a or 4T1 exosomes + sg-Tlr3 lentivirus. I Effect of IRF3 over-expression on IRF3, P-IRF3 and HAO1 expression in alveolar epithelial cells by Western blot analysis. J Diagram of the HAO1 promoter showing the location of IRF3 binding sites. K Luciferase activity of HAO1-promoter construct after transfection of IRF3 plasmid in alveolar epithelial cells. L Detection of IRF3, P-IRF3 and HAO1 expression in alveolar epithelial cells treated with 4T1 exosomes or 4T1 exosomes + shIRF3 by Western blot analysis. M Detection of oxalate production in alveolar epithelial cells treated with 4T1 exosomes or 4T1 exosomes + shIRF3. Means ± s.e.m are provided (n = 3). N Schematic diagram of the role of HAO1-mediated oxalate metabolism at pre-metastatic stage and metastatic stage. ***P 

Figure 4 STING Knockout Mitigates the Inflammatory Response in Diabetic Mice Subjected to I/R Injury. Mice were treated as described in Figure 2. (A) Representative western blotting results of p-TBK, p-IRF3, and IL-1β. (B) Statistical analysis of p-TBK/TBK in each group. (C) Statistical analysis of p-IRF3/IRF3 in each group. (D) Statistical analysis of IL-1β/GAPDH in each group. (E) Immunohistochemical results of TNF-α and IL-1β. Data are expressed as means ± SD, n=3.