Journal: bioRxiv

Article Title: Smoothened turnover regulated by Hedgehog signaling in Drosophila

doi: 10.64898/2026.01.08.698469

Figure Lengend Snippet: (A-C) Sagittal views of wing discs stained with (A,A’) α-Smo (gray), α-Ptc (green), and phalloidin (blue), or (B,B’) expressing BAC-GFP:Smo (green), BAC-Cherry:Smo (red), and mIFP ( ptc-Gal4>mIFP; Smo:GFP:Cherry/+ ); dashed lines indicate apical surface. (C-G) Wing disc expressing BAC-GFP:Smo (green) and Cad:Cherry; (C) sagittal, (D-G) frontal views in A compartment, P compartment and in apical or basolateral sections. (H-L) Same as (C-G), except wing discs express Cad:GFP (green) and BAC-encoded Cherry:Smo (red).

Article Snippet: The following antibodies were used: mouse α-GFP (Roche), rabbit α-RFP (Rockland), mouse α-Ptc (DSHB, Apa1), mouse α-En (DSHB, 4D9), DAPI, Phalloidin, mouse α-Dlg1, (DSHB, 4F3), α-Rab7 , α-Smo (DSHB, 20C6), Alexa633 conjugated-Phalloidin (Invitrogen), DAPI (Invitrogen), AbN ( ).

Techniques: Staining, Expressing

Journal: bioRxiv

Article Title: Smoothened turnover regulated by Hedgehog signaling in Drosophila

doi: 10.64898/2026.01.08.698469

Figure Lengend Snippet: Wild-type wing disc stained with α-Smo (red) and α-Ptc (green) at apical (A,A’) and basolateral (B,B’) optical planes. (A,B) Smo and Ptc staining; (A’,B’) Smo staining only. (A”,B”) Graphs showing intensity plots of images in (A,B) across A/P axis. (C,D) Same as (A,B) for unfixed disc expressing BAC-encoded GFP:Smo and mIFP driven by ptc-Gal4 showing fluorescence of Smo (green) and mIFP (blue) ( ptc-Gal4>mIFP ; GFP: Smo / + ). (E,F) Same as (C,D) for disc expressing BAC-encoded Cherry:Smo (red) and mIFP driven by ptc-Gal4 ( ptc-Gal4>mIFP ; Cherry : Smo / + ). (G) Same as (F) showing membrane-localized fluorescence only.

Article Snippet: The following antibodies were used: mouse α-GFP (Roche), rabbit α-RFP (Rockland), mouse α-Ptc (DSHB, Apa1), mouse α-En (DSHB, 4D9), DAPI, Phalloidin, mouse α-Dlg1, (DSHB, 4F3), α-Rab7 , α-Smo (DSHB, 20C6), Alexa633 conjugated-Phalloidin (Invitrogen), DAPI (Invitrogen), AbN ( ).

Techniques: Staining, Expressing, Fluorescence, Membrane

Journal: bioRxiv

Article Title: Smoothened turnover regulated by Hedgehog signaling in Drosophila

doi: 10.64898/2026.01.08.698469

Figure Lengend Snippet: (A-B’) Wing discs expressing mCD8:Cherry (red) by hh -Gal4 stained with α-Cut antibody to mark myoblasts (green). (A) notum region; (B) higher magnification image of boxed region in (A). Dashed white line encircles myoblasts. (C) Distribution of BAC encoded Hh:GFP (green) in the myoblasts (bounded by white dashed line; α-RFP, red). (D) Myoblasts encircled by white dashed line and marked by mCD8:Cherry (15B03Gal4>mCD8:Cherry; α-RFP, red) and α-Ptc antibody staining (green). (E-E”) Myoblasts (outlined with white dashed line and marked by expression of Cut, blue) expressing BAC encoded GFP:Cherry:Smo stained with α-GFP (green) and α-RFP (red) antibodies. (F) Clones induced as in expressing SmoRNAi (green) stained for Ptc (α-Ptc, red) bounded by white dashed line. Wild-type myoblasts are bounded by blue dashed line. (G-G”) Myoblasts (outlined with white dashed line and marked by expression of Mef2-Gal4>mIFP, blue) expressing BAC encoded GFP:Cherry:Smo (green) and α-RFP (red).

Article Snippet: The following antibodies were used: mouse α-GFP (Roche), rabbit α-RFP (Rockland), mouse α-Ptc (DSHB, Apa1), mouse α-En (DSHB, 4D9), DAPI, Phalloidin, mouse α-Dlg1, (DSHB, 4F3), α-Rab7 , α-Smo (DSHB, 20C6), Alexa633 conjugated-Phalloidin (Invitrogen), DAPI (Invitrogen), AbN ( ).

Techniques: Expressing, Staining, Clone Assay

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