Journal: Nature Communications

Article Title: High p16 expression and heterozygous RB1 loss are biomarkers for CDK4/6 inhibitor resistance in ER + breast cancer

doi: 10.1038/s41467-022-32828-6

Figure Lengend Snippet: A Half-maximal inhibitory concentration (IC 50 ) and IC 50 fold-change (fc) values for ribociclib, fulvestrant and the combination of drugs in T47D and MCF7 cells overexpressing p16 compared to control cells (MOCK), treated for 6 days and evaluated as shown underneath. Data are presented as mean values ± SEM of at least three independent experiments. B Immunoblot of the indicated proteins in MOCK and p16 overexpressing T47D and MCF7 cells untreated or treated with ribociclib for 5 days at the indicated concentrations. C Immunoblot of the indicated proteins in a cell competition assay. Cells were treated with vehicle or ribociclib 1 µM for the indicated days. D Comparison of structural models for the complexes of p16 bound to P18IN003 and p16 bound to CDK4. Gray cartoon is p16, orange spheres represent P18IN003, cyan cartoon is CDK4; zoomed view are the binding pocket with p16 shown as gray surface and highlighting the residues in the binding pocket of p16 as sticks and the hydrogen bonds made between P18IN003 and p16 shown as black dashed lines. E Relative spheroid area of PDC191 and PDC313 after treatment with 1 µM ribociclib, 20 nM P18IN003 and the combination for 7 days. Data are presented as mean values of three independent experiments ± SEM. Two-tailed p -values are based on the one-way ANOVA test with Tukey’s method correction compared with the vehicle (black line). Dashed line indicates the optimal cut-off established in Fig. . p16 and pRb scores of each PDC are indicated. F IC 50 and IC 50 fc values for ribociclib, fulvestrant and the combination of drugs in T47D and MCF7 cells overexpressing cyclin D1, compared to control cells (MOCK), treated for 6 days and evaluated as shown underneath. Data are presented as mean values ± SEM from at least three independent experiments. G Immunoblot of indicated proteins in control (MOCK) and cyclin D1-overexpressing T47D and MCF7 cells untreated or treated with 0.5 µM ribociclib for 48 h. H Immunoblot as indicated for panel ( C ). Source data are provided as a Source Data file.

Article Snippet: Fulvestrant and letrozole were purchased from Selleckchem.

Techniques: Concentration Assay, Control, Western Blot, Competitive Binding Assay, Comparison, Binding Assay, Two Tailed Test

Journal: Nature Communications

Article Title: High p16 expression and heterozygous RB1 loss are biomarkers for CDK4/6 inhibitor resistance in ER + breast cancer

doi: 10.1038/s41467-022-32828-6

Figure Lengend Snippet: A Waterfall plot showing the growth of n = 23 PDX treated with ribociclib 75 mg/kg plus alpelisib 35 mg/kg (bars and black dots) and vehicle (white circles). The percentage change from the initial volume is shown at day 35 of treatment. Dashed lines indicate the range of PD (>20%), SD (20% to −30%) and PR/CR (<−30%). The number of tumors treated per model is indicated in the brackets (n). Hashtags (#) indicate models harboring mutations in PIK3CA . Data represent mean values and error bars ± SEM. Boxes underneath show the molecular and intrinsic tumor subtype as well as their responses to the indicated treatments. The preclinical benefit to each drug is indicated as percentage in brackets. n.d, not determined. B IHC analysis of KI67 (left graph ; p -value < 0.001) and phospho-pRb S807/811 (right graph; p -value < 0.01) in vehicle and alpelisib plus ribociclib-treated PDXs according to the PDX response to alpelisib plus ribociclib. Data are presented as mean values of each PDX. Two-tailed p- values are based on Mann-Whitney U test for all biological replicates. Different colors represent the tumor’s intrinsic subtype. R, resistant; S, sensitive; Alp, alpelisib; Rib, ribociclib. C Relative tumor growth of PDX039 and PDX191 after treatment with the indicated drugs and time. Dashed lines indicate the range of PD (>1.2), SD (1.2 to −0.7) and PR/CR (<−0.7). Two-tailed p -values are based on the two-way ANOVA test with Bonferroni’s method correction. V, vehicle; R, ribociclib; A, alpelisib. D , E Relative tumor growth of the ribociclib-resistant PDX244LR#18R and PDX450 after treatment with the indicated drug(s) for the indicated period of time, respectively. Dashed lines indicate the range of PD (>1.2), SD (1.2 to −0.7) and PR/CR (<−0.7). F Immunoblot of the indicated proteins in PDC476.2 treated with vehicle or 500 nM palbociclib as single-agent or combined with 100 nM fulvestrant and/or 2.5 µM alpelisib in ex vivo cultures for 48 h. At least two independent experiments were conducted. Source data are provided as a Source Data file.

Article Snippet: Fulvestrant and letrozole were purchased from Selleckchem.

Techniques: Two Tailed Test, MANN-WHITNEY, Western Blot, Ex Vivo

Journal: Molecular imaging and biology : MIB : the official publication of the Academy of Molecular Imaging

Article Title: A Real-Time Non-invasive Auto-bioluminescent Urinary Bladder Cancer Xenograft Model

doi: 10.1007/s11307-016-0989-y

Figure Lengend Snippet: Development of auto-bioluminescent cells in vitro. a, c, e 0, 2.5, 5, 50, and 500 (×104) J82-Lux, J82-Ras-Lux, and SW780-Lux cells were plated in opaque 96-well plate and imaged with the IVIS Lumina system. b, d, f BLI (ph/s) signals were quantitated and analyzed. Columns, mean ± SEM of triplicates. g Lysates of J82-Ras and J82-Ras-Lux cells were prepared and analyzed by immunoblotting to detect levels of H-Ras, p-Mek, Mek, p-Erk, Erk, p-Akt, and Akt, with β-actin as a control. Data are representative of three independent experiments.

Article Snippet: Cell Culture and Reagents Human J82, SW780 (American Type Culture Collection [ATCC], Rockville, MD, USA), and oncogenic H-Ras(V12)-expressing, J82-Ras cells [ 6 ] were maintained in DMEM supplemented with 10 % FBS.

Techniques: In Vitro, Western Blot, Control

Journal: ACS Omega

Article Title: Glycerol-3-phosphate Acyltransferase1 Is a Model-Agnostic Node in Nonalcoholic Fatty Liver Disease: Implications for Drug Development and Precision Medicine

doi: 10.1021/acsomega.0c02350

Figure Lengend Snippet: Functional transcriptomic analysis. (A) Hepatic GPAT 1 mRNA expression level in the different groups #, p < 0.05 vs sham; *, p ≪ 0.01 vs sham. (B) Hepatic GPAT 1 mRNA expression exhibits a significant and direct correlation correlated with NAS and a logarithmic relation (C) with hepatic Picrosirius red staining.

Article Snippet: Blocking was performed with Background Sniper reagent (BS966M, Biocare Medical Pacheco, CA) for 11 min. A rabbit anti-GPAT1 antibody (NBP1-76907, Novus Biologicals, Centennial, CO) was diluted with 1% bovine serum albumin (BSA) in PBS + 0.3% Triton X-100 to a final concentration of 20 μg/mL and applied to each slide.

Techniques: Functional Assay, Expressing, Staining

Journal: ACS Omega

Article Title: Glycerol-3-phosphate Acyltransferase1 Is a Model-Agnostic Node in Nonalcoholic Fatty Liver Disease: Implications for Drug Development and Precision Medicine

doi: 10.1021/acsomega.0c02350

Figure Lengend Snippet: GPAT1 expression in NAFLD. GPAT1 immunofluorescence (red) is virtually absent in the sham liver (A, representative, 40×) but intense in the FFD liver (B, representative, 40×). Hepatocyte nuclei (white arrow) immunofluorescence green.

Article Snippet: Blocking was performed with Background Sniper reagent (BS966M, Biocare Medical Pacheco, CA) for 11 min. A rabbit anti-GPAT1 antibody (NBP1-76907, Novus Biologicals, Centennial, CO) was diluted with 1% bovine serum albumin (BSA) in PBS + 0.3% Triton X-100 to a final concentration of 20 μg/mL and applied to each slide.

Techniques: Expressing, Immunofluorescence