Figure 1.
2D gel map of tear protein A: Control. B: Dry eye syndrome.
Tear protein (30 µg) separated on 17 cm, pH 3–10 IPG strip in the first dimension and 13% SDS-PAGE in the second dimension. The differentially expressed proteins in DES compared to control are numbered.
Table 1.
Differentially expressed tear proteins identified by LC-MS/MS.
Table 2.
Down regulation of LPRR 4 protein in various types of DES cases as determined by 2D electrophoresis and PDQuest analysis.
Figure 2.
A representative DIGE image showing the tear protein profile.
8 peptide spots identified as LPRR4 that was down regulated in Dry eye condition are shown within the square box. DES case and control tear protein (30 µg) were labeled with Cy5 and Cy3 respectively as described in methods section. The range of the horizontal dimension is isoelectric point (from pI 3 to 10) using 17 cm IPG strips; the range of the vertical dimension is molecular weight (from approx. 97 to 3 kDa) on a 13% SDS-PAGE.
Figure 3.
3D DeCyder image of LPRR4 after DIGE.
Each protein in 3D view is shown. The 3D peak of each protein was generated based on the pixel intensity versus pixel area, normalized by the peak area of standard (Cy-2-labeled). A: control B: DES. DES showed a significant reduction of these five peptides identified as LPRR4 (p = 0.009) as observed by BVA analysis using Decyder software.
Table 3.
LPRR4 levels in tear from various types of DES by ELISA.
Figure 4.
Pearson’s Correlation graph of LPRR4 levels in Dry eye syndrome with clinical parameters namely Schirmer’s value and Tear Breakup Time (TBUT).
A. Tear LPRR4 levels vs Schirmer value (p = 0.008), B. Tear LPRR4 levels vs TBUT. (p = 0.005).
Figure 5.
Distribution graph of LPRR4 levels based on Dry eye syndrome (DES) grade, grade 1(mild DES) to grade 4 (severe DES).
A shift in the median was observed compared to the control.
Figure 6.
mRNA expression of LPRR4 in human lacrimal gland tissue as 144 bp product using RT-PCR showing tissue specificity for LPRR4.
Lane 1∶100 and 200 bp ladder, Lane 2: Negative control (except cDNA), Lane 3–6: human lacrimal gland tissue (2 µg), Lane 7–8: human corneal epithelial tissue ((2 µg).