Effects of S100A7 overexpression on the LPS-induced mRNA and protein expression of IL-6 and IL-8.

<p>(<b>A</b>) IL-6 and IL-8 mRNA levels and (<b>B</b>) protein levels were assayed after stable HaCaT cells (S100A7-EGFP or vehicle-EGFP) were exposed to LPS (100 or 1000 ng/ml, 24 h). The data represent the means ± SEM of three independent experiments. Statistical significance is indicated (*<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001). veh, vehicle; veh-100 or veh-1000, vehicle with 100 or 1000 ng/ml LPS treatment; pso, S100A7; pso-100 or pso-1000, S100A7-EGFP HaCaT cells with 100 or 1000 ng/ml LPS treatment.</p

Effects of S100A7 overexpression on mitochondrial dynamics and autophagy activation.

<p>S100A7-HaCaT cells were treated for 24 h with LPS at 100 and 1000 ng/ml. (<b>A</b>, <b>B</b>) The mitochondrial dynamics-related proteins Mfn1, Mfn2 and DRP1 and (<b>C</b>, <b>D</b>) the autophagy-related proteins Beclin1 and LC3B were detected by Western blot (<b>A</b>, <b>C</b> western blot images; <b>B</b>, <b>D</b> statistical results). Values are means ± SEM. Statistical significance is indicated (*<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001, ^#<i>P</i><0.05, ∧∧<i>P</i><0.01, ∧∧∧<i>P</i><0.001). veh, vehicle; veh-100 or veh-1000, vehicle with 100 or 1000 ng/ml LPS treatment; pso, S100A7; pso-100 or pso-1000, S100A7-EGFP HaCaT cells with 100 or 1000 ng/ml LPS treatment.</p

Characterization of S100A7 overexpression in HaCaT cells.

<p>(<b>A</b>) S100A7 mRNA levels in HaCaT cells overexpressing S100A7. S100A7 mRNA is normalized to β-actin. The data represent the mean ± SEM of three independent experiments. Statistical significance (**<i>P</i><0.01, ^##<i>P</i><0.01). (<b>B</b>) Immunofluorescence analysis of S100A7 overexpression. Cells were stained for EGFP (green) and anti-S100A7 (red); the nuclei were stained with DAPI (blue). Veh, vehicle; Veh-EGFP, vehicle-EGFP; pso-EGFP, S100A7-EGFP.</p

Effects of S100A7 overexpression on the expression of mitochondrial biogenesis and function.

<p>S100A7-EGFP and vehicle-EGFP HaCaT cells were exposed to lipopolysaccharide (LPS) (100 or 1000 ng/ml, 24 h). (<b>A</b>, <b>B</b>, <b>C</b>) The mRNA levels of PPAR-coactivator-1alpha (PGC-1α), nuclear respiratory factor-1 (NRF1) and transcription factor A (Tfam) were analyzed by quantitative RT-PCR. (<b>D</b>, <b>E</b>, <b>F</b>, <b>G</b>) Relative mitochondrial DNA contents (mtDNA), mitochondrial membrane potential (MMP), intracellular intracellular adenosine 5′-triphosphate (ATP) level and reactive oxygen species (ROS) generation were measured respectively. Values are means ± SEM; *Statistical significance (*<i>P</i><0.05, **<i>P</i><0.01). veh, vehicle; veh-100 or veh-1000, vehicle with 100 or 1000 ng/ml LPS treatment; pso, S100A7; pso-100 or pso-1000, S100A7-EGFP HaCaT cells with 100 or 1000 ng/ml LPS treatment.</p

Effects of S100A7 knockdown on MMP and mitochondrial homeostasis.

<p>(<b>A</b>) Expression of S100A7 mRNA after siRNA transfection. HaCaT cells were transfected with S100A7 siRNA using lipofectamin 2000 for 48 h. Total RNA was extracted and subjected to quantitative RT-PCR. The amount of S100A7 mRNA relative to β-actin is shown. (<b>B</b>) S100A7-siRNA HaCaT cells were treated for 24 h with LPS at 100 and 1000 ng/ml. The MMPs were assayed. (<b>C</b>, <b>D</b>) The mitochondrial dynamics-related proteins DRP1 and Mfn2 and the autophagy-related protein LC3B were detected by Western blot (<b>C</b>, Western blot images; <b>D</b>, statistical results). Values are means ± SEM. Statistical significance is indicated (^#<i>P</i><0.05, *<i>P</i><0.05, **<i>P</i><0.01, ***<i>P</i><0.001). cont, control; veh-si, vehicle-siRNA; veh-si-100 or veh-si-1000, vehicle-siRNA cells with 100 or 1000 ng/ml LPS treatment; pso-si, S100A7-siRNA; pso-si-100 or pso-si-1000, S100A7-siRNA HaCaT cells with 100 or 1000 ng/ml LPS treatment.</p