Journal: Science Advances

Article Title: A noncanonical function of the tyrosine degradation enzyme FAH drives CDK4/6 inhibitor resistance in breast cancer

doi: 10.1126/sciadv.aef1145

Figure Lengend Snippet: ( A ) Table with top 10 potential FAH interactors in MCF7 FAH-NLS BioID cells treated with 200 nM abemaciclib. Table shows average peptide counts from triplicate samples analyzed by mass spectrometry. CTRL BioID and FAH-NES BioID peptide counts are shown as controls. ( B ) Bar chart of GO molecular function pathway enrichment of proteins interacting with FAH-NLS under abemaciclib treatment. Data are plotted as fold enrichment of GO terms that are overrepresented in the dataset compared to the background set and FDR of the enrichment. snRNA, small nuclear RNA. ( C and D ) Representative images (C) and quantification (D) of nuclear PLA foci in vehicle- and abemaciclib-treated MCF7 FAH-V5 cells. PLA was used to assess FAH (V5-tag) and CDK9 interaction in 100 cells. ( E and F ) Enrichment score of MYC targets and B-MYB targets in MCF7 EV and FAH-OE cells (E) and T47D gAAVS and gFAH#3 KO cells (F) treated with abemaciclib for 72 hours. Yu MYC targets up and Shepard BMYB targets through the mSigDB were used for the analysis. ( G to I ) Western blot analysis of pRB, cMYC, FAH, and vinculin (VCL) protein in vehicle- and abemaciclib-treated MCF7 EV control and FAH -expressing cells (G), patient #26 EV and FAH PDOs (H) and T47D gAAVS and gFAH #3 KO cells (I). Biological replicates: n = 2 (D). Red line indicates median. Mann-Whitney test was used to assess significance. Scale bar, 10 μm.

Article Snippet: The membranes were blocked with 5% nonfat milk in 0.1% Tween 20 in tris-buffered saline for 1 hour and incubated with primary antibodies for 1 hour in room temperature or overnight at 4°C using the following antibodies: FAH (Novus, NBP2-48736), c-MYC/N-MYC (CST, #13987), pRB (CST, #9307), and VCL (CST, #13901).

Techniques: Mass Spectrometry, Analysis, Western Blot, Control, Expressing, MANN-WHITNEY

Journal: Science Advances

Article Title: A noncanonical function of the tyrosine degradation enzyme FAH drives CDK4/6 inhibitor resistance in breast cancer

doi: 10.1126/sciadv.aef1145

Figure Lengend Snippet: ( A ) AlphaFold prediction of FAH-CDK9 interaction. AlphaFold multimer prediction was performed using the Cosmic 2 platform. ( B ) Quantification of nuclear FAH-D143K in MCF7 cells treated with vehicle or 200 nm abemaciclib for 72 hours. ( C and D ) Representative images (C) and quantification (D) of PLA foci in vehicle- and abemaciclib-treated MCF7 FAH-WT-V5 and FAH D143K-V5 cells. PLA was used to assess V5-tag and CDK9 interaction in 100 cells. ( E ) Abemaciclib IC 50 of MCF7 EV, FAH-WT, and FAH D143K cells treated with increasing doses of abemaciclib for 72 hours. PrestoBlue cell viability reagent was used to assess cell viability. ( F ) Western blot analysis of pRB, cMYC, FAH, and vinculin (VCL) protein in vehicle- and abemaciclib-treated MCF7 EV control, FAH-WT, and FAH-D143K cells. ( G ) Representative confocal images of cleaved caspase 3 and EdU staining, ( H ) quantification of EdU+ cells, and ( I ) cleaved caspase 3+ PDOs in patient #26 PDOs treated with vehicle, 300 nM abemaciclib, 15 nM AZD4573 (CDK9 inhibitor), or the combination of abemaciclib and AZD4573 for 96 hours. Proliferation was assessed by pulsing PDOs with EdU for 4 hours and quantifying the ratio of EdU+ cells per total (DAPI+) number of cells. Apoptosis was quantified by staining for cleaved caspase 3–positive PDOs. Ten to 15 PDOs were analyzed per each individual experiment. Biological replicates: n = 3 (D), n = 5 (E), and n = 3 [(H) and (I)]. Error bars indicate mean + SD, and red line indicates median. Student’s unpaired t test [(H) and (I)], one-way ANOVA with Dunnetts’ test (E), or Kruskal-Wallis with Dunn’s test (D) was used to assess significance. Scale bars, 10 μm (C) and 40 μm (H).

Article Snippet: The membranes were blocked with 5% nonfat milk in 0.1% Tween 20 in tris-buffered saline for 1 hour and incubated with primary antibodies for 1 hour in room temperature or overnight at 4°C using the following antibodies: FAH (Novus, NBP2-48736), c-MYC/N-MYC (CST, #13987), pRB (CST, #9307), and VCL (CST, #13901).

Techniques: Western Blot, Analysis, Control, Confocal, Staining

Journal: bioRxiv

Article Title: Cellular source and circuit context organize functional specificity in the Drosophila NPF system

doi: 10.64898/2026.09.08.750118

Figure Lengend Snippet: Key Resources Table

Article Snippet: Antibody , Rabbit anti-NPF , Ray Biotech, Inc , RB-19-0001 , IHC.

Techniques: Imaging, Electron Microscopy, Saline, Drosophila, Protease Inhibitor, Yeast, Formulation, Sample Throughput, Feeding Assay, Microscopy, Software, Analysis