Journal: Shock (Augusta, Ga.)

Article Title: TARGETING S100A9-TLR2 AXIS CONTROLS MACROPHAGE NLRP3 INFLAMMASOME ACTIVATION IN FATTY LIVER ISCHEMIA REPERFUSION INJURY

doi: 10.1097/SHK.0000000000002470

Figure Lengend Snippet: S100A9 induces inflammatory injury via macrophage TLR2 activation BMMs were isolated from WT mice and incubated with rS100A9 or rS100A8 respectively for 24 h. (A) Western-assisted analysis of TLR2 and ATF4 in rS100A9-stressed macrophages. (B) Western-assisted analysis of TLR2 and ATF4 in rS100A8-stressed macrophages; BMMs were transfected with the TLR2-siRNA or control vector followed by incubation with rS100A9. (C) Detection of cytokines IL-1β, CXCL1, CXCL-2, and IL-6 by qRT-PCR in BMMs. (D) Western-assisted analysis of ATF4 in nuclear extracts. (E) Immunofluorescence staining for ATF4 distribution in macrophages. DAPI was used to visualize nuclei. Scale bars: 50 μm, 20 μm. N = 3–6/group. All data represent the mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001. BBM, bone-derived macrophage. WT, wild type; qRT-PCR, quantitative real-time PCR.

Article Snippet: After a period of 24–48 h, the cells were treated with recombinant S100A9 (rS100A9) or rS100A8 (Prospec, Rehovot, Israel) for additional 24 h.

Techniques: Activation Assay, Isolation, Incubation, Western Blot, Transfection, Control, Plasmid Preparation, Quantitative RT-PCR, Immunofluorescence, Staining, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Aging (Albany NY)

Article Title: S100A8 and S100A9, both transcriptionally regulated by PU.1, promote epithelial-mesenchymal transformation (EMT) and invasive growth of dermal keratinocytes during scar formation post burn

doi: 10.18632/aging.203112

Figure Lengend Snippet: S100A8 and S100A9 were upregulated in post burn skin and thermal-stimulated keratinocytes. ( A ) Representative blots for S100A8 and S100A9 in burned and matched normal skin tissues from burn patients. Comparison of ( B ) S100A8 and ( C ) S100A9 in burned and matched normal skin tissues of 38 patients. ( D ) Expression of S100A8 and S100A9 and ( F ) EMT marker proteins in thermal-stimulated keratinocytes was detected with Western blotting at different time points. ( E ) Double immunofluorescence assay for cellular localization of S100A8 and S100A9 and ( G ) immunofluorescence assay for nuclear translocation of Snail protein in thermal-stimulated keratinocytes at different time points. *** P < 0.001.

Article Snippet: Human recombinant S100A8 and S100A9 proteins (rS100A8 and rS100A9) were purchased form Abnova Corporation (Chinese Taipei).

Techniques: Comparison, Expressing, Marker, Western Blot, Immunofluorescence, Translocation Assay

Journal: Aging (Albany NY)

Article Title: S100A8 and S100A9, both transcriptionally regulated by PU.1, promote epithelial-mesenchymal transformation (EMT) and invasive growth of dermal keratinocytes during scar formation post burn

doi: 10.18632/aging.203112

Figure Lengend Snippet: S100A8 and S100A9 promote cell proliferation, expression of EMT marker proteins, and cell invasion in primary keratinocytes. Human primary keratinocytes were treated with 1 μg of rS100A8, 1 μg of rS100A9, 0.5 μg of rS100A8 plus 0.5 μg of rS100A9, or 1 μg of rS100A8 plus 1 μg of rS100A9, heat-inactivated rS100A8/rS100A9 were applied as a negative control. After incubation for 48 h, ( A ) cell number, ( B ) expression levels of EMT marker proteins, ( C ) proportion of CD44+CD24- cells, and ( D ) cell invasion, were respectively detected with automatic cell counter, Western blotting, FACS, and Transwell migration assay. * P < 0.05 vs. control, # P < 0.05 vs 1 μg rS100A8, $ P < 0.05 vs. 0.5 μg rS100A8 + 0.5 μg rS100A9.

Article Snippet: Human recombinant S100A8 and S100A9 proteins (rS100A8 and rS100A9) were purchased form Abnova Corporation (Chinese Taipei).

Techniques: Expressing, Marker, Negative Control, Incubation, Western Blot, Transwell Migration Assay, Control

Journal: Aging (Albany NY)

Article Title: S100A8 and S100A9, both transcriptionally regulated by PU.1, promote epithelial-mesenchymal transformation (EMT) and invasive growth of dermal keratinocytes during scar formation post burn

doi: 10.18632/aging.203112

Figure Lengend Snippet: PU.1 was positive associated with expression of S100A8/9 and identified as a transcription factor of them. ( A ) Transcriptionally activity of PU.1 on ( A ) S100A8 and ( B ) S100A9 gene promoters was detected by luciferase reporter gene assay. Pearson correlation coefficient assay was used to evaluate the correlations of ( C ) S100A8 and S100A9 expression levels, ( D ) S100A8 and PU.1 expression levels, and ( E ) S100A9 and PU.1 expression levels. ChIP-qPCR assay was used to validate the binding of PU.1 with ( F ) S100A8 promoter and ( G ) S100A9 promoter. ** P < 0.01.

Article Snippet: Human recombinant S100A8 and S100A9 proteins (rS100A8 and rS100A9) were purchased form Abnova Corporation (Chinese Taipei).

Techniques: Expressing, Activity Assay, Luciferase, Reporter Gene Assay, ChIP-qPCR, Binding Assay

Journal: Aging (Albany NY)

Article Title: S100A8 and S100A9, both transcriptionally regulated by PU.1, promote epithelial-mesenchymal transformation (EMT) and invasive growth of dermal keratinocytes during scar formation post burn

doi: 10.18632/aging.203112

Figure Lengend Snippet: S100A8 and S100A9 promoted scar development in PU.1 -/- burn model mice. ( A ) expression of PU.1, S100A8 and S100A9 proteins in the skin of WT and skin-specific PU.1 knockout (PU.1 -/- or ΔPU.1) mouse. The effect of overexpression of S100A8 or/and S100A9 on ( B ) expression of EMT marker proteins, ( C ) time of wound healing, ( D ) histopathological changes (by HE staining), ( E ) scar volume, ( F ) scar thickness, and ( G ) contents of collagens in PU.1 -/- burn model mice. * P < 0.05 vs. WT, # P < 0.05 vs. ΔPU.1+Vector.

Article Snippet: Human recombinant S100A8 and S100A9 proteins (rS100A8 and rS100A9) were purchased form Abnova Corporation (Chinese Taipei).

Techniques: Expressing, Knock-Out, Over Expression, Marker, Staining, Plasmid Preparation