Figure 2. PLOD3 mRNA and protein expression in normal brain tissue and human glioma cell lines.

(A, B) Expression of PLOD3 mRNA (A) and protein in LN229, GBM8401 and U118MG glioma cell lines (B) and normal brain tissue. The quantitative results of PLOD3 mRNA expression represent data from three independent experiments. The relative gene expression was standardized with that in normal brain. ^*p < 0.05 and ^***p < 0.001 compared to the normal brain tissue group. (C) Lysates were collected from GBM8401 and LN229 cells 48 h after transfection with 25 nM siRNA. Blots were probed with antibodies targeting total PLOD3. Duplicate biological samples were collected for GBM8401 and LN229 cells. β-actin was used as a loading control. (D, E) GBM8401 and LN229 cells were transfected with 25 nM siRNA. Cell number was determined at the indicated time points. The data are expressed as the mean ± s.d.; n = 3; ^**p < 0.01, and ^***p < 0.001. (F, G) GBM8401 and LN229 cells were stained with propidium iodide (PI) for cell cycle analysis using flow cytometry. The data are expressed as the mean ± s.d.; n = 3; ^*p < 0.05, ^**p < 0.01. (H, I) GBM8401 and LN229 cells were labeled with BrdU and then processed for flow cytometric analysis. The data are shown as the means ± s.d.; n = 3; ^*p < 0.05. (J, K) GBM8401 and LN229 cells were labeled with C₁₂FDG for senescence activity assay by flow cytometric method. The data are shown as the mean ± s.d.; n = 3; ^*p < 0.05, and ^***p < 0.001 compared to the siPLOD3 group. (L) Western blot analysis was performed to detect changes in the expression of autophagy associated proteins. The results represent data from two independent experiments. ACTN served as a loading control.