Journal: Oncology Letters

Article Title: High S phase kinase-associated protein 2 expression is a potential prognostic biomarker for glioma

doi: 10.3892/ol.2020.11818

Figure Lengend Snippet: Correlation of SKP2 expression with other biomarkers in 395 glioma tissues. (A) Blue and brown staining represents negative and positive expression of p-Rb, respectively. (B) According to linear correlation analysis, SKP2 was positively correlated with p-Rb in both HGG and LGG groups. ‘r’ means the coefficient of correlation. (C) Blue and brown staining represents negative and positive expression of EGFR, respectively. (D) Positive immunostaining was regarded as high EGFR expression. And no immunostaining was regarded as low EGFR expression. Mann-Whitney U test was used to evaluate the association between SKP2 and EGFR. SKP2, S phase kinase-associated protein 2; EGFR, epidermal growth factor receptor; p-Rb, phosphorylated retinoblastoma protein; HGG, high-grade glioma; LGG, low-grade glioma.

Article Snippet: The primary SKP2 antibody (1:100; cat. no. 2652S; CST Biological Reagents Co., Ltd.), anti-isocitrate dehydrogenase 1 (IDH1 R132H ) antibody (working solution; cat. no. ZM0447; OriGene Technologies, Inc.), phosphorylated retinoblastoma protein (p-Rb) antibody (1:100; cat. no. 8516T, CST Biological Reagents Co., Ltd.), epidermal growth factor receptor (EGFR) antibody (working solution; cat. no. ZA-0505; OriGene Technologies, Inc.) were added and the sections were incubated at 4°C overnight.

Techniques: Expressing, Staining, Immunostaining, MANN-WHITNEY

Journal: Frontiers in Genetics

Article Title: Assembly factor for spindle microtubules ( ASPM ) promotes osimertinib resistance in lung cancer by increasing EGFR stability

doi: 10.3389/fgene.2025.1593314

Figure Lengend Snippet: ASPM enhances EGFR resistance in lung cancer cells. (A) Dose‒effect relationship curves of osimertinib for PC-9 cells; (B) Dose‒effect relationship curves of osimertinib for PC-9 OR cells. (C) mRNA expression levels of ASPM and EGFR in PC-9 and PC-9 OR cells. (D) Protein expression of ASPM and EGFR in PC-9 and PC-9 OR cells. The histogram represents the relative gray values of the intracellular EGFR and ASPM proteins, and the data are presented as the means ± standard deviations (means ± SDs, n = 3 fields). (E) Silencing of the ASPM gene and mRNA expression levels of ASPM in PC-9 and PC-9 OR cells. (F) Dose‒effect relationship curves of osimertinib in PC-9 si ASPM cells. (G) Dose‒effect relationship curves of osimertinib in PC-9 OR si ASPM cells. (H) mRNA expression levels of ASPM were detected in PC-9 cells after overexpressing the ASPM gene. (I) Dose‒effect curves of osimertinib after PC-9 overexpression of ASPM . (J) The viability of the cells in each group was assessed via a CCK-8 assay. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The membrane was blocked with 5% skim milk for 60 min and incubated with primary antibodies, including ASPM (Proteintech, 26223-1-AP) (1:600) and EGFR (Cusabio, CSB-PA005571LA01HU) (1:1000), overnight at 4 °C, followed by 2 h of incubation with HRP-conjugated goat anti-rabbit (Proteintech, SA00001-2) (1:2000) at room temperature.

Techniques: Expressing, Over Expression, CCK-8 Assay

Journal: Frontiers in Genetics

Article Title: Assembly factor for spindle microtubules ( ASPM ) promotes osimertinib resistance in lung cancer by increasing EGFR stability

doi: 10.3389/fgene.2025.1593314

Figure Lengend Snippet: ASPM plays a key role in the stabilization of drug-resistant cell lines. (A) Scatter plot of the correlation between ASPM and EGFR expression; p < 0.05. (B) Silencing of ASPM followed by ASPM and EGFR mRNA expression. (C) Protein expression of EGFR after silencing ASPM. (D) CHX administration at various time points. The right panel represents the relative gray values of the intracellular EGFR proteins, and the data are presented as the means ± standard deviations (means ± SDs, n = 3 fields). (E) IF images of PC-9 and PC-9 OR cells incubated with primary antibodies against EGFR (red) and ASPM (green). Detection was performed using secondary antibodies coupled to Alexa Fluor 488 and Alexa Fluor 555. The cell nuclei were stained with DAPI (blue), Scale = 7.5 μm. (F) mRNA expression levels of ASPM in PC-9 OR cells after silencing of ASPM -2. (G) PC-9 OR cells in the NC group/si ASPM -2 group were stained with propidium iodide (PI), and the DNA content was detected via flow cytometry. The red color represents the theoretical curve that we fit via ModFit software on the basis of the DNA content distribution data of the cells. (H) Statistics of the flow cytometric results in H plots. n = 3, a p value <0.05 was considered statistically significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The membrane was blocked with 5% skim milk for 60 min and incubated with primary antibodies, including ASPM (Proteintech, 26223-1-AP) (1:600) and EGFR (Cusabio, CSB-PA005571LA01HU) (1:1000), overnight at 4 °C, followed by 2 h of incubation with HRP-conjugated goat anti-rabbit (Proteintech, SA00001-2) (1:2000) at room temperature.

Techniques: Expressing, Incubation, Staining, Flow Cytometry, Software

Journal: Frontiers in Genetics

Article Title: Assembly factor for spindle microtubules ( ASPM ) promotes osimertinib resistance in lung cancer by increasing EGFR stability

doi: 10.3389/fgene.2025.1593314

Figure Lengend Snippet: ASPM is significantly upregulated in NSCLC tumor tissues and strongly associated with reduced patient survival. ASPM silencing attenuates PC-9 and PC-9 OR malignant phenotypes, including proliferation and invasion, and sensitizes resistant cells to osimertinib. In addition, inhibiting the expression of ASPM effectively reduces damage to the cell cycle and protein stability of drug-resistant cells, thereby restoring the expression and function of EGFR.

Article Snippet: The membrane was blocked with 5% skim milk for 60 min and incubated with primary antibodies, including ASPM (Proteintech, 26223-1-AP) (1:600) and EGFR (Cusabio, CSB-PA005571LA01HU) (1:1000), overnight at 4 °C, followed by 2 h of incubation with HRP-conjugated goat anti-rabbit (Proteintech, SA00001-2) (1:2000) at room temperature.

Techniques: Expressing