Journal: The Journal of Biological Chemistry

Article Title: The role of DNAJC3 in enhancing glioma progression and regulating the tumor immune microenvironment

doi: 10.1016/j.jbc.2025.111059

Figure Lengend Snippet: DNAJC3 promotes the survival of glioma cells by activating the MAPK–ERK and PI3K–AKT signaling pathways. A , enrichment analysis of GSEA. B , WB was used to detect the effects of silencing or overexpression of DNAJC3 on the phosphorylation levels of ERK and AKT (n = 3, Student's t test, compared with si-NC/vector). The data are presented as mean ± SD. (∗∗∗ p < 0.001; ∗∗ p < 0.01; ∗ p < 0.05; DNAJC3, DNAJC3 overexpression group; si-DNAJC3, DNAJC3 knockdown group; si-NC, negative control of the knockdown group; vector, empty vector control of the overexpression group.) DNAJC3, DnaJ heat shock protein family member C3; ERK, extracellular signal–regulated kinase; GSEA, gene set enrichment analysis; MAPK, mitogen-activated protein kinase; WB, Western blot.

Article Snippet: After blocking the membrane for 1 h in Tris-buffered saline with Tween-20 containing 5% nonfat milk, the membrane was incubated overnight at 4 °C with primary antibodies specific for DNAJC3 (1:1000 dilution, 26721-1-AP; Proteintech), GAPDH (1:2000 dilution, 60004-1-Ig; Proteintech), GRP78 (1:2000 dilution, 66574-1-Ig; Proteintech), AKT (1:1000 dilution, 10176-2-AP; Proteintech), p-AKT (1:1000 dilution, 66444-1-Ig; Proteintech), ERK (1:1000 dilution, 67170-1-Ig; Proteintech), and p-ERK (1:1000 dilution, 28733-1-AP; Proteintech).

Techniques: Protein-Protein interactions, Over Expression, Phospho-proteomics, Plasmid Preparation, Knockdown, Negative Control, Control, Western Blot