Journal: Scientific Reports

Article Title: Discovery of a Proteolysis Targeting Chimera for TRKA and RET-derived oncoproteins

doi: 10.1038/s41598-025-25687-w

Figure Lengend Snippet: Pz-1-based PROTACs activity on CCDC6-RET. TPC1 cells were treated with the indicated PROTACs or Pz-1 for 12 h at the indicated concentrations. Cells were harvested, and extracts were subjected to Western blotting with anti-RET antibody. Tubulin was used as a loading control.

Article Snippet: Antibodies for VEGFR2 (2479), phospho-TRKA (Tyr490) (9141), phospho-TRKA (Tyr674/675) (4621), panTRK (A7H6R) (92991) and RET (14698) were from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Activity Assay, Western Blot, Control

Journal: Scientific Reports

Article Title: Discovery of a Proteolysis Targeting Chimera for TRKA and RET-derived oncoproteins

doi: 10.1038/s41598-025-25687-w

Figure Lengend Snippet: Compound 9 activity on RET- and TRKA-driven cell proliferation. The indicated cell lines were incubated with vehicle (NT: not treated) or increasing concentrations of compound 9 and counted at the indicated time points. Data are the mean ± SD of a single experiment performed in triplicate. Number of cells plated at Day 0: KM12 (10,000/well); TT (200,000/well); TPC-1 (5000/well); MZ-CRC-1 and Lc-2/ad (100,000/well). IC 50 dose was calculated using PRIZM software (GraphPad Software Inc). Confidence intervals are indicated in brackets.

Article Snippet: Antibodies for VEGFR2 (2479), phospho-TRKA (Tyr490) (9141), phospho-TRKA (Tyr674/675) (4621), panTRK (A7H6R) (92991) and RET (14698) were from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Activity Assay, Incubation, Software

Journal: Molecular Oncology

Article Title: Dual targeting of RET and SRC synergizes in RET fusion‐positive cancer cells

doi: 10.1002/1878-0261.70155

Figure Lengend Snippet: Signaling analysis of pralsetinib and dasatinib combination effects in RET + non‐small cell lung cancer (NSCLC) cells. (A) (Left) Western blot analysis of indicated signaling proteins in CUTO32 cells after treatment with pralsetinib (1 μ m ), dasatinib (0.1 μ m ) or their combination for 3 h. (Right) Quantification of pRET ( n = 4), pSRC ( n = 3), pPAK1/2 ( n = 4), pERK1/2 ( n = 3), pAKT ( n = 3), and pS6 ( n = 3) blots, normalized by loading controls (Actin or vinculin). (B) (Left) Western blot analysis of indicated signaling proteins after treatment with pralsetinib (1 μ m ), dasatinib (0.1 μ m ) or their combination for 3 h in CUTO32 cells expressing SRC wild‐type (WT) or SRC T341I gatekeeper. (Right) Quantification of pRET ( n = 3), pSRC ( n = 3), pPAK1/2 ( n = 4), pERK1/2 ( n = 3), pAKT ( n = 3), and pS6 ( n = 3) blots, normalized by loading controls (Actin or vinculin). Statistical significance was determined by one‐way ANOVA with Tukey's multiple comparisons; n.s., not significant. Results are shown as mean ± SD.

Article Snippet: Primary antibodies against PARP1 (9542), cleaved Caspase‐3 (9661), p27 (2552), p21 (2947), p‐RET (Y905, 3221), RET (3223) p‐SRC (Y419, 2101), SRC (2108), p‐PAK1/2 (S144/S141, 2606), PAK1/2/3 (2604), p‐ERK1/2 (Y204/Y187, 4370), p‐AKT (S473, 9271), AKT (9272), p‐S6 (S235/S236, 4858), S6 (2217), and Vinculin (13901) were acquired from Cell Signaling Technology.

Techniques: Western Blot, Expressing

Journal: Molecular Oncology

Article Title: Dual targeting of RET and SRC synergizes in RET fusion‐positive cancer cells

doi: 10.1002/1878-0261.70155

Figure Lengend Snippet: Combination effects of the next‐generation SRC tyrosine kinase inhibitor (TKI) eCF506 with pralsetinib in RET + non‐small cell lung cancer (NSCLC) cells. (A) (Left) Western blotting of indicated signaling proteins in CUTO32 cells after treatment with pralsetinib (1 μ m ), eCF506 (0.1 μ m ) or their combination for 3 h. (Right) Quantification of p‐RET ( n = 3), p‐SRC ( n = 3), pPAK1/2 ( n = 3), pERK1/2 ( n = 3), pAKT ( n = 3), and p‐S6 ( n = 3) blots, normalized by loading controls (Actin or vinculin). (B) (Top) Cell viability of CUTO32 ( n = 3) and LC‐2/Ad ( n = 3) cells upon treatment with RET TKIs alone and in combination with eCF506 (0.1 μ m ) for 3 days. (Bottom) Cell viability and Bliss synergy analysis of RET + NSCLC cells at single concentrations of RET TKIs (0.625 μ m , see gray arrows above) and eCF506 (0.1 μ m ). (C) Clonogenic survival, quantification and Bliss synergy analysis of RET + NSCLC cells ( n = 4 for CUTO32 and n = 3 for LC‐2/Ad) after treatment with pralsetinib (1 μ m ), eCF506 (0.1 μ m ) or their combination for 7 days. (D) Western blotting and quantification of apoptosis markers upon treatment of CUTO32 cells with pralsetinib (1 μ m ), eCF506 (0.1 μ m ) and their combination for 3 days ( n = 3). Statistical significance was determined by one‐way ANOVA with Tukey's multiple comparisons; n.s., not significant. Results are shown as mean ± SD.

Article Snippet: Primary antibodies against PARP1 (9542), cleaved Caspase‐3 (9661), p27 (2552), p21 (2947), p‐RET (Y905, 3221), RET (3223) p‐SRC (Y419, 2101), SRC (2108), p‐PAK1/2 (S144/S141, 2606), PAK1/2/3 (2604), p‐ERK1/2 (Y204/Y187, 4370), p‐AKT (S473, 9271), AKT (9272), p‐S6 (S235/S236, 4858), S6 (2217), and Vinculin (13901) were acquired from Cell Signaling Technology.

Techniques: Western Blot