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. 2024 Feb 2;15:988. doi: 10.1038/s41467-024-45283-2

Fig. 4. Chronic precursor trapping induces the formation of TNTs.

Fig. 4

a Representative confocal images showing HeLaGAL cells over-producing mCherry (red) and Su9-EGFP-DHFR in the presence of 100 nM MTX for 48 h (+ MTS + MTX; chronic trapping). Arrows indicate protrusions and the box highlights zoom region. b Quantification of (a) demonstrating the proportion of transfected cells with protrusions. N = 4 biological replicates; 20 cells counted per replicate. P = 0.9886, <0.0001, 0.9505, 0.7149, 0.0004, >0.9999. c Representative confocal images showing tubulin-α and β-actin staining of protrusions following chronic trapping (48 h) in HeLaGAL cells. N = 4 biological replicates. d Quantification of the proportion of HeLaGAL cells, subjected to chronic precursor trapping (48 h), with protrusions following incubation for 48 h in the absence (vehicle; DMSO only) or presence of 100 nM nocodazole (48 h). N = 3 biological replicates; 20 cells counted per replicate. P = 0.0004. Statistical significance was determined using a two-way ANOVA and Šidák’s test (b) or a two-tailed, unpaired t test (d). Data are presented as mean values ± S.D. with individual data points for each biological replicate and P values are reported left-right, bottom-top (b, d). Raw data are provided in the Source Data file (b, d). MTS mitochondrial targeting sequence, MTX methotrexate, TNT tunnelling nanotube.