Fig. 2

LDHA is highly expressed and succinylated at K222 in GC. a The gene expression of LDHA in GC samples (red) and paired normal tissues (green). Data from the GEPIA website. b Analysis of mRNA levels of LDHA in GC and adjacent normal tissues by qRT-PCR. c Analysis of the protein levels of LDHA in GC and adjacent normal tissues by western blot. d The lactate levels in GC and adjacent normal tissues. e Identification of succinylated LDHA peptides by mass spectrometry analysis of GC tissues. f K222 is the main succinylation site of LDHA. Cells (293 T) were transfected with the indicated plasmids. Then, immunoprecipitation (IP) was performed, followed by western blot analysis of succinylation. g Analysis of K222suc-LDHA levels in GC and adjacent normal tissues using two K222 succinylation-specific antibodies. h and i Immunohistochemical analysis of LDHA (h) and K222suc-LDHA (i) in GC and adjacent normal tissues. j The K222 site in the three-dimensional structure of the LDHA homotetramer. Data from the RCSB PBD, http://www.rcsb.org/3d-view/6MV8). Data are presented as mean ± SEM, *P < 0.05, ***P < 0.001