An Exorbitant RhoC Conspriracy

In the current study, we have shown for the first time that cultured IPE cells express functional TLRs and respond to PAMPs through activation of TLRs, particularly TLR2, TLR3 and TLR4. This study extends the current knowledge of the role of TLR activation in iris culture and uveal innate immune mechanism in the pathogenesis of ocular inflammation. Conclusion Our results demonstrate that IPE cells express functional TLRs and their activation may have implications for the pathogenesis of ocular inflammation. Abbreviations IPE: Iris pigment epithelial cells; RPE: Retinal pigment epithelial cells; PAMPs: Pathogen-associated molecular patterns; TLRs: Toll-like receptors; LPS: Lipopolysaccharides; GSK1210151A MALP-2: Macrophage buy INCB024360 activating lipopeptide-2. Competing interests Authors declare no conflict of interest. Authors' contributions KM designed, performed and analysed all the experiments and prepared the manuscript. JJYC assisted in data analyses, presentation and helped to draft the manuscript. NDG, PJM and DW designed experiments, supervised all aspects of the project and revised the manuscript. All authors read and approved the final manuscript. Supplementary Material Additional file 1: Human IPE and RPE stained for cytokeratin. The primary IPE and RPE, cultured from iris and retina, of human donors, were probed with anti-pan cytokeratin antibody (1:200 dilution; A) and an isotype control antibody (mouse IgG1; 1: 200 dilution). Immunohistochemistry was performed to confirm the epithelial origin of primary IPE and RPE. A representative of cytokeratin stained IPE was shown in A and no staining was seen in the negative control (IgG1). The result was confirmed by flow cytometry (B). The peak representing cytokeratin stained cells (in pink) was shifted away from that of the isotype control antibody (in grey). Click here for file(89K, pdf) Additional file 2: Expression of TLR transcripts and proteins in human IPE and RPE. Human IPE (lane 1) and RPE (lane 2) from the same donor were cultured to confluence and expression of TLR1 to TLR10 genes and proteins was investigated by reverse transcription PCR (A) and Western RhoC blotting (B) using specific human TLR1 to TLR10 primers and antibodies, respectively. M= 100 bp DNA ladder (100, 200, 300, 400, 500, up to 1000 bp from bottom to top). TLR mRNA expression was measured by densitometry and normalised against GAPDH which served as a loading control. Normalised TLR mRNA expression levels are presented as mean �� SD (N=3) (C). Two-way ANOVA and Bonferroni��s multiple comparison test were used to analyse the data, *p