Scheme 1. Overview of the Workflow of MicroPOTS for the Identification of Proteomic Changes in ∼200 CP-A Cells after Treatment with Either LCA or X-ray.

First, cells were grown in keratinocyte media, and the generated CP-A null cells and wild-type were subjected to different stressors as previously described. (a) Samples were then processed for protein extraction and further digested into peptides using the microPOTS system. (b) The collected peptides were subsequently subjected to LC-MS/MS analysis. (c) A spectrum showing the relative intensity and mass to charge ratio (m/z) of the ions being analyzed. (d) The resulting files were loaded into MaxQuant for peptide identification, after which the output files of this step were next imported into R environment and analyzed using the DEP package. (e) The results of the analysis were then visualized using the previously mentioned R packages.