Fig. 6: The intracellular domain of NOTCH1 interacts with SMAD2/3 in C2C12 cells.
From: Molecular mechanism of Activin receptor inhibition by DLK1
A Illustration of in situ proximity ligation assay (PLA), with antibodies targeting SMAD2/3 and NOTCH1. PLA uses secondary antibodies with oligonucleotides that can form a rolling circle amplification when both probes are in close proximity. Created in BioRender. Antfolk, D. (2025) https://BioRender.com/cyz9ujyB Immunofluorescence images showing the association between SMAD2/3 and NOTCH1 detected by in situ PLA. Untreated C2C12 cells (control), Myostatin-treated cells, and Myostatin + soluble DLK1 (sDLK1) treated cells were analyzed. Actin cytoskeleton stained by phalloidin 488. Nuclei counterstained by Hoechst 33342 (blue). Scale bar, 50 μm. The experiment was independently repeated three times. C Dot plot depicts quantification of manually counted PLA dots per cell +/− SEM from the experiments performed in Fig. 6B. The total number of cells counted per treatment, n = 53 (Control), n = 65 (Myostatin), and n = 55 (sDLK1 + Myostatin). Statistics were obtained using a one-way ANOVA in Prism 10 (Version 10.4.0) with Tukey’s multiple comparisons post hoc test (p < 0.0001, 95% CI [−33.05, −25.35] for Control vs Myostatin; p = 0.1869 (ns), 95% CI [−6.985, 1.027] for Control vs soluble DLK1 + Myostatin; and p < 0.0001, 95% CI [22.41, 30.04] for Myostatin vs soluble DLK1 + Myostatin). Source data are provided as a Source Data file.
