Fig 1: PLCD3 regulates the RAP1 pathway through PKC. A. Mountain plot analysis of co-expression correlation of PLCD3 and PKC in lung cancer tumor tissues. B. Immunoblot detection of PLCD3, PKC, Rapl, b-Raf in lung cancer cells with or without PLCD3 silencing and PKC restoration. C-F. Analysis of GLUT1, HK1, HK2 and PKM2 in PLCD3-silenced and PLCD3-restored lung cancer cells. All data were shown as the mean ± SD, n = 3, *p < 0.05, **p < 0.01.
Fig 2: PLCD3 promotes aerobic glycolysis and cell growth through activation of the RAP1 pathway. A. KEGG shows significant enrichment of RAR1 pathway-related genes. B. Western blot analysis of Rap1, b-Raf, GLUT1, HK1, HK2, and PKM2 in lung cancer cells with or without PLCD3 silencing. C. Growth assay of A549 cells with and without PLCD3 induction and XIV induction. D–E. Glucose uptake and consumption assays in A549 and NCI-H1299 cells with or without PLCD3 induction and 1 μmol/L GGTI298 (Rap1 inhibitor) treatment. F-I. Detection of the mRNA expression levels of GLUT1, HK1, HK2 and PKM2 in A549 and NCI-H1299 cells transfected or not transfected with PLCD3 and GGTI298. (F) GLUT1; (G) HK1; (H) HK2; (I) PKM2. J. The effect of PLCD3 and GGTI298 on the invasion of lung cancer cells was detected by transwell assay. All data were shown as the mean ± SD, n = 3, *p < 0.05, **p < 0.01.
Fig 3: Western blot analysis of B-RAF, K-RAS, and C-RAF protein expression. (A) Western blot analysis for HCT-116 and (B) Western blot analysis for HT-29 colon cancer cells. The proteins are represented in the upper, middle, and lower panels, respectively. Cells were treated with MSN, nanoformulation, and free ELG at their respective IC50 concentrations and incubated for 48 h. Lane 1: Untreated control; Lane 2: MSN treatment; Lane 3: Nanoformulation treatment; Lane 4: ELG treatment. B-actin was used as loading control (last panel).
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