patched 1 antibody  (Affinity Biosciences)

Journal: bioRxiv

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancer

doi: 10.1101/2025.09.03.673759

Figure Lengend Snippet: A . Relative PTCH1 RNA expression in TNBC (red), Her2 (blue), luminal B (LB, green), and luminal A (LA, orange) in the TCGA cohort is illustrated by box plots (log 2 transformed). Outliers are shown within each population (open circles). Student’s t test was used to compare RNA levels between two groups. The p values are indicated (*P < 0.05; **P < 0.01; ***P < 0.001; ns P > 0.05). B . Normalized PTCH1 mRNA expression according to TNBC (IHC) status from all DNA microarray data from bc-GenExMiner v4.5 illustrated by box plots (log 2 transformed) (Jezequel et al 2021). C . Boxplots showing PTCH1 mRNA expression (normalized counts from RNA seq data) in circulating tumor cells (CTC) isolated from metastasis breast cancer patient peripheral blood (Wurth R et al 2025). D . High PTCH1 mRNA expression is associated with a poorer prognosis in breast cancer. Distant Metastasis Free Survival (DMFS) and disease-free survival (DFS) data based on PTCH1 mRNA expression were obtained using the exhaustive prognostic analysis on all status breast cancers on the bc-GenExMiner v4.5 web portal, and illustrated by Kaplan–Meier (KM) curves for all ER and PR breast cancers with positive nodules. The obtained Hazard Ratio (HR) with 95% confidence interval and log-rank p-values are shown.

Article Snippet: After 1 h at room temperature in blocking buffer (20 mmol/L Tris-HCl pH 7.5, 45 mmol/L NaCl, 0.1 % Tween-20, and 5 % non-fat milk), nitrocellulose membranes were incubated overnight at 4 °C with the monoclonal rat anti-PTCH1 antibody from R&D system biotechne clone 413220 (1/1000), the monoclonal mouse anti-P-gp antibody from abcam (ab3366, 1/1000), the rabbit anti-ABCG2 antibody from GeneTex (GTX50793, 1/500) or the rabbit anti-GAPDH antibody from Elabscience (1/20000).

Techniques: RNA Expression, Transformation Assay, Expressing, Microarray, RNA Sequencing, Isolation

Journal: bioRxiv

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancer

doi: 10.1101/2025.09.03.673759

Figure Lengend Snippet: Distant Metastasis Free Survival (DMFS), disease-free survival (DFS) and overall survival (OS) data based on PTCH1 mRNA expression were obtained from the intrinsic molecular subtypes’ prognostic analysis on ER+/HER2-high proliferative breast cancers performed on bc-GenExMiner v4.5 web portal and illustrated by Kaplan–Meier curves. The obtained Hazard Ratio (HR) with 95% confidence interval and log-rank P-values are shown.

Article Snippet: After 1 h at room temperature in blocking buffer (20 mmol/L Tris-HCl pH 7.5, 45 mmol/L NaCl, 0.1 % Tween-20, and 5 % non-fat milk), nitrocellulose membranes were incubated overnight at 4 °C with the monoclonal rat anti-PTCH1 antibody from R&D system biotechne clone 413220 (1/1000), the monoclonal mouse anti-P-gp antibody from abcam (ab3366, 1/1000), the rabbit anti-ABCG2 antibody from GeneTex (GTX50793, 1/500) or the rabbit anti-GAPDH antibody from Elabscience (1/20000).

Techniques: Expressing

Journal: bioRxiv

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancer

doi: 10.1101/2025.09.03.673759

Figure Lengend Snippet: A . PTCH1 mRNA expression (log 2 transformed) in various luminal and HER2 breast cancer cell lines (dark green) and in TNBC cell lines (other colors). TNBC cell lines are depicted according to the “Lehmann TNBC subtype” nomenclature : basal-like 1 (yellow), basal-like 2 (pale green), immunomodulatory (brown), luminal androgen receptor (dark pink), mesenchymal (pale pink) and mesenchymal stem-like (pink). B . PTCH1 protein expression in three TNBC cell lines. Western-blot was performed on 50 µg extracts from TNBC cell lines (MDA-MB-231, HCC-38 and MDA-MB-468) with antibodies directed against PTCH1. PTCH1 and GAPDH signals were quantified using ImageJ software. Data presented are the mean ± SEM of at least 3 independent experiments. Significance is calculated using Oneway ANOVA Turkey’s multiple comparisons test and attained at P < 0.05 (*).

Article Snippet: After 1 h at room temperature in blocking buffer (20 mmol/L Tris-HCl pH 7.5, 45 mmol/L NaCl, 0.1 % Tween-20, and 5 % non-fat milk), nitrocellulose membranes were incubated overnight at 4 °C with the monoclonal rat anti-PTCH1 antibody from R&D system biotechne clone 413220 (1/1000), the monoclonal mouse anti-P-gp antibody from abcam (ab3366, 1/1000), the rabbit anti-ABCG2 antibody from GeneTex (GTX50793, 1/500) or the rabbit anti-GAPDH antibody from Elabscience (1/20000).

Techniques: Expressing, Transformation Assay, Western Blot, Software

Journal: bioRxiv

Article Title: Inhibition of PTCH1 drug efflux activity enhances chemotherapy efficacy against triple negative breast cancer

doi: 10.1101/2025.09.03.673759

Figure Lengend Snippet:

Article Snippet: After 1 h at room temperature in blocking buffer (20 mmol/L Tris-HCl pH 7.5, 45 mmol/L NaCl, 0.1 % Tween-20, and 5 % non-fat milk), nitrocellulose membranes were incubated overnight at 4 °C with the monoclonal rat anti-PTCH1 antibody from R&D system biotechne clone 413220 (1/1000), the monoclonal mouse anti-P-gp antibody from abcam (ab3366, 1/1000), the rabbit anti-ABCG2 antibody from GeneTex (GTX50793, 1/500) or the rabbit anti-GAPDH antibody from Elabscience (1/20000).

Techniques: Concentration Assay

Journal: Nature communications

Article Title: Multiplatform molecular profiling uncovers two subgroups of malignant peripheral nerve sheath tumors with distinct therapeutic vulnerabilities.

doi: 10.1038/s41467-023-38432-6

Figure Lengend Snippet: Fig. 5 | SHH pathway is important for malignant transformation in a subset of MPNSTs and a therapeutic target. a Capillary-based immunoassay (WES) con- firming knockout of PTCH1 and subsequent increased GLI1 activity in HSC1-λ cells. b Trypan blue counts of parental HSC1-λ and PTCH1-knockout clone. Error bars, s.e.m.; n = 3 biologically independent experiments. c PTCH1-knockout in HSC1-λ cell lines leads to upregulation of SHH pathway (GLI1, GLI2 and SMO). Error bars, s.e.m.; n = 3 biologically independent experiments. d Protein blot confirming knockout of PTCH1 in ipNF06.2A cells. e Trypan blue counts of a parental ipNF06.2A cells and PTCH1-knockout clone. Error bars, s.e.m; n = 3 biologically independent experiments. f PTCH1-knockout in ipNF06.2A leads to upregulation of SHH pathway (GLI1, GLI2 and SMO). Error bars, s.e.m; n = 3 biologically indepen- dent experiments. g Representative images and quantitative cell migration in HSC1-

Article Snippet: Primary antibodies for B-Actin(1:1000, Cat #8457S, Cell Signaling Technologies), Vinculin(1:30000, Cat #V9264, Sigma Aldrich), PTCH1(1:500,Cat#MAB41051, R&D systems) and APC(1:500, Cat #15270, Abcam) were used.

Techniques: Transformation Assay, Knock-Out, Activity Assay, Migration