Journal: Molecules

Article Title: Exploring EGFR, Nectin-4, and TROP-2 as Therapeutic Targets for Bladder Cancer Photoimmunotherapy

doi: 10.3390/molecules30244802

Figure Lengend Snippet: ( a ) Western blot analysis of EGFR, Nectin-4, and TROP-2 expression in the BC cell lines RT4 and RT112 and in the antigen-negative control cell line CHO. ( b ) Cell binding of the antibodies and conjugates to the target cells as demonstrated by flow cytometry. Binding of the isotype control antibody is shown in gray.

Article Snippet: Following transfer, EGFR, Nectin-4 and TROP-2 were detected using polyclonal rabbit anti-human EGFR IgG (#sc-03-G, Santa Cruz Biotechnology, Dallas, TX, USA), polyclonal rabbit anti-human Nectin-4 IgG (#17402S, Cell Signaling Technologies, Danvers, MA, USA), and monoclonal rabbit anti-human TROP-2 IgG (#90540, Cell Signaling Technologies).

Techniques: Western Blot, Expressing, Negative Control, Binding Assay, Flow Cytometry, Control

Journal: Molecules

Article Title: Exploring EGFR, Nectin-4, and TROP-2 as Therapeutic Targets for Bladder Cancer Photoimmunotherapy

doi: 10.3390/molecules30244802

Figure Lengend Snippet: Graphical abstract illustrating the future clinical application of PIT using WB692-CB2 conjugates for the treatment of NMIBC. Following intravesical application of the antibody–dye conjugates into the bladder, they can be activated by cystoscopic red light for the selective eradication of BC cells expressing EGFR, Nectin-4 and/or TROP-2. For personalized PIT, a pre-therapeutic determination of target antigen expression in biopsy samples may guide the selection of an optimal conjugate combination. Created in BioRender. Wolf, I (2025) https://BioRender.com/sx866ld .

Article Snippet: Following transfer, EGFR, Nectin-4 and TROP-2 were detected using polyclonal rabbit anti-human EGFR IgG (#sc-03-G, Santa Cruz Biotechnology, Dallas, TX, USA), polyclonal rabbit anti-human Nectin-4 IgG (#17402S, Cell Signaling Technologies, Danvers, MA, USA), and monoclonal rabbit anti-human TROP-2 IgG (#90540, Cell Signaling Technologies).

Techniques: Expressing, Selection

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Sorcin regulates alveolarization and airway tissue remodeling during lung morphogenesis

doi: 10.1007/s00018-025-05870-y

Figure Lengend Snippet: Altered protein expression and EGFR signaling in Sorcin-deficient lung development A ) Confocal microscopy of 3-week-old lung tissue from WT and Sri⁻ / ⁻ mice. Sri⁻ / ⁻ lung tissue shows reduced SP-B (red) expression compared to WT. Scale bars, 20 μm B ) Western blot analysis of lung lysates from 3-week-old WT and Sri⁻ / ⁻ mice. Sri⁻ / ⁻ mice exhibit decreased EGFR, PANRAS and RAB5C protein levels compared to WT. Densitometry quantification is shown (mean ± SEM). ** p < 0.01 (Student’s t-test), *** p < 0.001 (determined by Student’s t-test) C-E ) Confocal microscopy of 3-week-old lung sections: WT mice show epithelial localization of EGFR (green) (C), RAB5C (green) (D) and PANRAS (red) protein (E). In Sri⁻ / ⁻ mice, these proteins are significantly reduced. Scale bars, 20 μm

Article Snippet: Then, sections were blocked in 5% goat serum for 1 h and incubated overnight at 4 °C with primary antibody rabbit polyclonal EPCAM/CD326 (1:100 in 1%BSA-PBS solution) (Proteintech, #21050-1-AP), rabbit polyclonal SFTPB (1:100 in 1%BSA-PBS solution) (Thermo Fisher Scientific, # PA542000 ), rabbit polyclonal E-cadherin (1:100 in 1%BSA-PBS solution) (Proteintech, #20874-1-AP), mouse monoclonal E-cadherin (1:100 in 1%BSA-PBS solution) (BD Bioscience, #610181), mouse monoclonal α-Smooth Muscle Actin (1:100 in 1%BSA-PBS solution) (Sigma-Aldrich, #A5228), rabbit polyclonal EGFR (1:100 in 1%BSA-PBS solution) (Proteintech, #30847-1-AP) rabbit polyclonal RAB5C (1:100 in 1%BSA-PBS solution) (Thermo Fisher Scientific, #PA5101828), mouse monoclonal PANRAS (Ab3) (1:100 in 1%BSA-PBS solution) (Sigma-Aldrich, #OP40) followed by incubation with Alexa fluor 488 (rabbit)-conjugated secondary antibodies (1:500 in 1%BSA-PBS solution) (Thermo Fisher Scientific).

Techniques: Expressing, Confocal Microscopy, Western Blot

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