Journal: bioRxiv

Article Title: The combination of BCL-xL PROTAC and mTOR inhibitor sensitizes pancreatic ductal adenocarcinoma to KRAS G12D inhibitor treatment by enhancing apoptosis induction

doi: 10.64898/2026.01.05.697773

Figure Lengend Snippet: Immunoblot analyses of phosphorylated- and total- ERK, AKT and S6, BIM extra-long isoform (BIM EL ) and NOXA in AsPC1 (a), HPAC (b), SW1990 (c), and PANC1 cells (d) after they were treated with indicated concentrations of MRTX1133 for 24 h. The β-tubulin was used as an equal loading control. e-h. Densitometric analysis of BIM EL and NOXA immunoblots in AsPC1 (e), HPAC (f), SW1990 (g), and PANC1 cells (h). Immunoblots detect three isoforms of BIM i.e., short isoform (BIM S ), long isoform (BIM L ) and extra-long isoform (BIM EL ). Among them, BIM EL is the major isoform and is shown here. i-l. mRNA expression levels of BIM-coding gene BCL2L11 in AsPC1 (i), HPAC (j), SW1990 (k), and PANC1 cells (l) after 24 h treatment with MRTX1133. m-p . mRNA expression levels of NOXA-coding gene PMAIP1 in AsPC1 (m), HPAC (n), SW1990 (o), and PANC1 cells (p) after 24 h treatment with MRTX1133. Data in i-p are presented as mean ± SD (n= 3 biological replicates). ** p <0.01, *** p <0.001, **** p <0.0001, ns p >0.05 i.e., not significant compared to untreated (control) cells as determined by one-way ANOVA and Dunnett’s multiple comparisons test. Since ERK1 and ERK2 have very close molecular weights (∼44 and ∼42 kDa), the two bands appeared as a single merged band in panels a-d (and subsequent figures) due to limited resolution in gradient gels that we have used.

Article Snippet: The mRNA expressions were quantified using pre-designed Taqman probes of BCL2L11 (Assay #Hs00708019_s1) and PMAIP1 (Assay # Hs00560402_m1) from Thermo Fisher.

Techniques: Western Blot, Control, Expressing

Journal: Pharmaceutics

Article Title: Pharmacodynamic-Driven Sequence-Dependent Synergy Effects in Pemetrexed-Osimertinib Combination Against Non-Small Cell Lung Cancer (NSCLC): Optimizing Synergy Through Sequential Interval

doi: 10.3390/pharmaceutics17081044

Figure Lengend Snippet: The pharmacodynamic interaction between PEM-induced apoptotic signaling and OSI-induced pro-apoptotic protein Bim under different sequencing conditions . ( A ) HCC827 cells were treated with 1 μM PEM for 72 h, and the apoptosis-associated proteins were monitored by WB ( n = 4). ( B ) HCC827 cells were treated with 20 nM OSI for 72 h, and the apoptosis-associated proteins were monitored by WB ( n = 4). ( C ) Schematic illustration of the pharmacodynamic interactions between PEM-induced apoptosis signal and OSI-induced BIM upregulation in different sequencing conditions. ( D ) PEM did not show synergy when co-exposed with 5 μM ABT263 for 72 h but shows strong synergy at PEM 72 h → 5 μM ABT 263 regimens ( n = 6). ( E , F ) Apoptosis-associated proteins and apoptosis rate of HCC827 following indicated treatments (the concentrations of PEM and ABT263 were 1 μM and 5 μM, respectively) ( n = 3). ( G ) PEM shows synergy at PEM 72 h → 20 μM BIM BH3 72 h regimens (transfected by protein transfection agent) ( n = 6). ( H , I ) Apoptosis rate and apoptosis-associated proteins of HCC827 following indicated treatments (the concentrations of PEM and BIM BH3 were 1 μM and 34.1 μM, respectively) ( n = 3). ( J ) Schematic of sequential interval modulation for optimal synergy in the PEM → OSI regimen. All data were presented as mean ± SD, * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. NEG (negative control): drug-free treatment with freshly seeded cells at high viability; BLK (blank control): drug-free medium or vehicle treatment. All corresponding WB images for each panel in this figure were shown in .

Article Snippet: Bim BH3, a 20-residue peptide containing the BH3 domain from the BH3-only protein Bim, was purchased from MCE (Monmouth Junction, NJ, USA, Catalog HY-P1527).

Techniques: Sequencing, Transfection, Negative Control, Control