Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Role of hydroxymethylglutharyl-coenzyme A reductase in the induction of stem-like states in breast cancer

doi: 10.1007/s00432-024-05607-7

Figure Lengend Snippet: Expression of HMGCR is variable and response to statins subtype-dependent in BC. HMGCR expression was analyzed by RT-qPCR in Luminal (MC-7, T47D, BT474) and triple-negative (MDA-MB-468, HCC70, MDA-MB-231, Hs578T) BC cell lines. Hepatocellular carcinoma hepG2 cell line was used as positive control and the immortalized non-transformed mammary epithelial line MCF-10A as negative control. Data were analyzed by one-way ANOVA, followed by Dunnett’s multiple comparison test (vs MCF-10A) (a) . Viability of MCF-7, T47D, MDA-MB-231 and Hs578T cell lines after 48 h treatment with SIM, LOVA or vehicle (Control). Data were analyzed by one-way ANOVA, followed by Dunnett’s multiple comparison test (vs Control; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001) (b) . Representative images of MCF-7, T47D, MDA-MB-231 and Hs578T cell lines treated with 10 µM SIM, 10 µM LOVA or vehicle (Control). Scale bar: 100 µm (c) . SIM: Simvastatin; LOVA: Lovastatin

Article Snippet: BC human cell lines MCF-10A (ATCC Cat# CRL-10317, RRID:CVCL_0598), MCF-7 (ATCC Cat# HTB-22, RRID:CVCL_0031) and Hs578T (ATCC Cat# CRL-7849, RRID:CVCL_0332) were a gift from Dr. Luthy’s lab, and T47D (ATCC Cat# HTB-133, RRID:CVCL_0553), BT474 (ATCC Cat# CRL-7913, RRID:CVCL_0179), MDA-MB-231 (ATCC Cat# CRL-12532, RRID:CVCL_0062), MDA-MB-468 (ATCC Cat# HTB-132, RRID:CVCL_0419) and HCC70 (ATCC Cat# CRL-2315, RRID:CVCL_1270) were a gift from Dr. Elizalde’s lab. BT474 were authenticated by the Human Cell Line Authentication STR Profiling Service from the Johns Hopkins University with the GenePrint 10 System (Promega).

Techniques: Expressing, Quantitative RT-PCR, Positive Control, Transformation Assay, Negative Control, Comparison, Control

Journal: Cell chemical biology

Article Title: A highly potent bi-thiazole inhibitor of LOX rewires collagen architecture and enhances chemoresponse in triple-negative breast cancer.

doi: 10.1016/j.chembiol.2024.06.012

Figure Lengend Snippet: Figure 4. Effects of LXG6403 on collagen crosslinking/deposition, drug penetration in 3D culture, and ROS/FAK/DNA damage axis (See also Figure S4) (A and B) Collagen I/fibronectin staining (A) and insoluble collagen assay (B) in ECM derived from HFF-1 cells treated with vehicle or 15 mM of LXG6403 (n = 2). Scale bar = 50 mm. (C and D) Collagen I/fibronectin staining (C) and insoluble collagen assay (D) in decellularized ECM (dECM) incubated with HCC1143 cells treated with 10 mM of 1 or LXG6403 (n = 2). Scale bar = 50 mm. (E–G) Relative doxorubicin autofluorescence in MDA-MB-231 (E) (n = 3), HCC1143 (F) (n = 3), cells and TNBC organoids (G) (n = 2 for vehicle and LXG6403, and n = 4 for doxorubicin and doxorubicin+LXG6403) embedded in collagen I and treated with the LOX inhibitor. (H) Relative mitochondrial ROS levels in MDA-MB-231 cells treated with cisplatin with or without LXG6403 (n = 4). (I) Western blot analysis in MDA-MB-231 cells treated with cisplatin or doxorubicin with or without LXG6403. Actin was used as the loading control. Data rep- resents mean values ± SD. p values were calculated with the unpaired, two-tailed Student’s t test. (*p < 0.05; **p < 0.01).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER BAPN Santa Cruz Cat#sc-214124 Cisplatin Medchemexpress Cat#HY-17394 Doxorubicin Medchemexpress Cat#HY-15142A Paclitaxel Medchemexpress Cat#HY-B0015 Matrigel Corning Cat#356231 Collagen-I Corning Cat#354236 Collagenase type III Worthington Cat#CLS-3 Cadaverine dihydrochloride Millipore Sigma Cat#33220 Putrescine dihydrochloride Thermo Scientific Cat#A18312-14 LOX (expression host: HEK293T) Origene Cat#TP313323 LOXL1 (expression host: Baculovirus infected Sf9 cells) Rockland Cat#009-001-S40 LOXL2 (expression host: HEK293) AcroBiosystem Cat#LO2-H52H3 Critical commercial assays Lysyl Oxidase Activity Assay (Cell-based) Abcam Cat#ab112139 Lysyl Oxidase Activity Assay (Recombinant protein-based) Abcam Cat#ab284573 Amplex Red Hydrogen Peroxide/Peroxidase Assay Kit Invitrogen Cat#A22188 MitoSOX Red Mitochondrial Superoxide Indicator, for live-cell imaging Invitrogen Cat#M36008 PicroSirius Red Abcam Cat#ab150681 DCFDA/H2DCFDA - Cellular ROS Assay Kit Abcam Cat#ab113851 Total Collagen Assay Abcam Cat#ab222942 Deposited data Raw western blot and microscopy images This paper Mendeley Data: https://doi.org/10.17632/ dzfhhfh3d8.1 Mass spectroscopy generated raw files This paper MassIVE Repository: MSV000094776/https:// massive.ucsd.edu/ProteoSAFe/static/massive.jsp Mass spectroscopy imaging data This paper PRIDE Repository: PXD052642/ https://www.ebi.ac.uk/pride/ Experimental models: Cell lines MDA-MB-231 ATCC Cat#HTB-26 HCC1937 ATCC Cat#CRL-2336 Hs-578-T ATCC CatHTB-126 HCC1143 ATCC Cat#CRL-2321 MCF-12A ATCC Cat#CRL-3598 HUVEC ATCC Cat#CRL-1730 HFF-1 ATCC Cat#SCRC-1041 Oligonucleotides siAllstar Qiagen Cat#1027280 siLOX Dharmacon Cat#D-009810-17-0005 sgcontrol ABM Cat#K010 Recombinant DNA hLOX-His Genescript, custom order N/A hLOXL2-His Genescript, custom order N/A wt LOX ORF Vector Builder, custom order N/A hLOX-HA Vector Builder, custom order N/A hLOXL2-HA Vector Builder, custom order N/A Software and algorithms Image Lab 3.0 BioRad N/A Inkscape 1.2.2 Inkscape Project N/A Graphpad Prism 8.0 Dotmatics N/A (Continued on next page) e2 Cell Chemical Biology 31, 1–16.e1–e11, November 21, 2024

Techniques: Staining, Collagen Assay, Derivative Assay, Incubation, Western Blot, Control, Two Tailed Test