Figure S2: PMTRIPs FISH-PLA is specific, effective, and sensitive without dextran sulfate. from Quantification and Localization of Protein–RNA Interactions in Patient-Derived Archival Tumor Tissue

Abstract

<p>Supplementary Figure 2: PMTRIPs FISH-PLA is specific, effective, and sensitive without dextran sulfate. A). 5 nM of V5-tagged PMTRIPs targeting polyadenylated mRNA were delivered as FISH probes, with FISH hybridization buffer containing either 0%, 5% or 10% dextran sulfate. PLA (white) was performed between the V5 tag and HuR. Extended focus images are shown. Scale bars indicate 10 µm. B) Quantification of PLA is shown. Statistics were performed with a one-way ANOVA with a Dunn's multiple comparisons test, where n=30 and **** p < 0.0001. 95% confidence intervals are shown in red. C) 5 nM V5 tagged MTRIPs targeting polyadenylated mRNA were assembled using either PNA or 2'OM oligos and were delivered as FISH probes. PLA (white) was performed between the V5 tag and HuR. Extended focus images are shown. Scale bars indicate 10 µm. Quantification of PLA is shown. Statistics were performed with a one-way ANOVA with a Dunn's multiple comparisons test, where n=30 and ** p < 0.002 and **** p < 0.0001. 95% confidence intervals are shown in black. D) Representative extended focus control images for figure 2A are shown. PLA is in white. Scale bars indicate 10 µm. E) Representative extended focus control images for figure 2B are shown. PLA is in white. Scale bars indicate 10 µm. F) Representative extended focus control images for figure 2C are shown. PLA is in white. Scale bars indicate 10 µm. G) Representative extended focus control images for figure 2D are shown. PLA is in white. Scale bars indicate 10 µm.</p>