circ-Hdgfrp3 interacts with p-bodies in murine MNs and in living mammalian cells. (A) Upper panels: two examples of colocalization between circ-Hdgfrp3 (magenta) and DCP1A (green) in nonstressed mESCs-derived MNs. Lower panels: two examples of association between circ-Hdgfrp3 (magenta), DCP1A (green), and TIAR (gray) in stressed mESCs-derived MNs. Oxidative stress response was monitored with TIAR (gray) staining. Magnified yellow boxes highlight the colocalizing spots. Scale bars, 10 μm. (B) Quantification of circ-Hdgfrp3 spots colocalizing with DCP1A signals in murine MNs in nonstressed (left panel) and stressed (right panel) conditions (NWT MNs = 25 cells; NWT MNs+NaAsO2 = 30 cells). (C) Representative HEK 293T cells expressing circ-HDGFRP3 labeled with HBC620 (magenta) and GFP-DCP1A (green). Lower panel shows a magnification view of the white box with circ-HDGFRP3 and DCP1A interacting over time (duration 15 min; interval ∼11 sec). Spots and tracks detected with TrackMate Plug-in on Fiji-ImageJ are overlaid, respectively, as magenta (circ-HDGFRP3) and green (DCP1A) circles and lines. Scale bars, 5 μm. (D) Single-particle tracking of circ-HDGFRP3 and DCP1A spots shown in C over time. The plot indicates circ-HDGFRP3 (magenta) and DCP1A (green) xy-coordinates (micron) as a function of time (min). (E) Scatter interval plot representing PCC between circ-HDGFRP3 and DCP1A over time upon NaAsO2 administration. Dots represent individual measurements of PCC in an interval of 5 min collected from 17 time-lapse acquisitions (duration 1 h; interval 1–5 min) from two independent biological replicates. Horizontal lines indicate mean values and error bars represent ± SEM.
