MRM validation of immune protein upregulation in Gipie-silenced ACC co-cultures (IMAGE)
Caption
Targeted phase proteomics by multiple reaction monitoring showing upregulation of immune activated proteins in immune cells in Gipie-silenced ACC -immune co-culture model.
(A) Granzyme A, (C) CD48, (E) Granzyme B, (G) HLA class I, antigen B, (I) HLA class I, antigen A; normalised relative protein expression graphs done with Graph Pad prism 9.4.1 showing t-test (Wilcoxon matched-pairs signed rank test) comparison data of immune proteins of NK cells from UM-HACC-2A co-culture model compared to NK cells from Gipie-silenced UM-HACC-2A co-culture model. ∗P≤ 0.05, ∗∗∗P≤ 0.0001. n = 24 biological replicates at different time points. (B, D, F, H, J). MRM ion chromatogram peaks of analytes. All peaks: Gaussian smooth width 0, retention time half window 30 s, noise percentage 40 %, baseline sub window 2 min, and peak splitting 2 points. (B) Granzyme A, +2, GVTSFGLENK. All three transitions (y8, y6, y5) shown. Expected retention time 32.70 min, (D) CD48, +2, LDPQSGALYISK. All three transitions (y10, y9, y8) shown. Expected retention time 32.70 min. (F) Granzyme B, +2, GDSGGPLVCNK. All three transitions (y8, y7, y6) shown. Expected retention time 36.49 min. (H) HLA class I, antigen B, +2, AYLEGECVEWLR. All three transitions (y9, y8, y6) shown. Expected retention time 47.60 min, (J) HLA class I, antigen A, +2, FIAVGYVDDTQFVR. All three transitions (y10, y8, y7) shown. Expected retention time 48.85 min.
Credit
Rajdeep Chakraborty, Thiri Zaw.
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