Increased Expression of Long Noncoding RNA LOC100506314 in T cells from Patients with Nonsegmental Vitiligo and Its Contribution to Vitiligo Pathogenesis

Figure 4

Validation of the proteins that interacted with LOC100506314. RNA pull-down assay was performed with the biotinylated LOC100506314. The biotin-labeled LOC100506314 was prepared in vitro by pcDNA3.1LOC100506314 using T7 RNA polymerase in the presence of the kit for RNA biotin labeling. An empty vector of pcDNA3.1 was used as a control. The extracted nuclear proteins were incubated with the biotin-conjugated LOC100506314 or control, followed by a pull-down with streptavidin-conjugated beads. The precipitated proteins were subjected to analysis by shotgun proteomics analysis. We chose three proteins, including (a) Signal transducer and activator of transcription 3 (STAT3), (b) Protein kinase R (PKR), and (c) macrophage migration inhibitory factor (MIF) that related to the immune response for further validation by western blotting.

(a)

(b)

(c)