Fig. 5
ZNF322A mediates heat shock protein 27 phosphorylation and elicits unfolded protein response. a The protein expressions of eIF2α in UPR signaling pathway were analysis by immnoblotting. b Phosphorylation of eIF2αS51 was determined and normalized to the level of total eIF2α. The normalized values were then compared between siControl and siZNF322A. c Enriched consensus motifs of MAPKAPK2 on HSP27S82 were predicted and identified in ZNF322A-silenced phosphoproteome. MS/MS spectrum for HSP27 (QIpSSGVSEIR), and fragment ions in the MS/MS spectrum localize at serine82 in HSP27. d Validation of HSPB1S82 phosphorylation using immunoblot analysis. e Phosphorylation of HSP27 was determined and normalized to the level of total HSP27. The normalized values were compared between siControl and siZNF322A groups. f Schematic representation of the heat stress responses of HSP27S82 via UPR pathway by silencing of ZNF322A in A549 lung cancer. β-actin is the internal control to normalize protein expression. The bars represent densitometric analysis of three biological replicates and the data are shown as mean ± SD. * p < 0.05