Fig. 4.

Measurement of distances between proteins of the MICOS complex. (A) Representative 3D two-color MINFLUX acquisition of Mic10 (CF680, blue) and Mic60 (Alexa Fluor 647, orange) in Mic10-TO HeLa cells. (Scale bar, 500 nm.) (B) Representative 3D two-color MINFLUX acquisition of Mic19 (CF680, blue) and Mic60 (Alexa Fluor 647, orange) in Mic10-TO HeLa cells. (Scale bar, 500 nm.) (C) Histograms of interspecies nearest-neighbor distances between Mic10 and Mic60 for a selection of MINFLUX acquisitions. A Gaussian fit (dashed line) determines Gaussian mean distance $d$ and spread of distances $\sigma$. (D) Same as in C, but now for Mic19 and Mic60. (E) Gaussian mean of the nearest-neighbor distances measuring the average distance between Mic10/Mic19 and Mic60 molecules for all acquisitions for Mic10–Mic60 (purple, n = 11) and Mic19–Mic60 (green, n = 10) with 25 and 75% percentiles (blue box), the median value (red line and number), extreme values (error bar), and outliers (red crosses). A Mann–Whitney U significance test delivered a probability of P for the mean distances to result from the same distribution. (F) Same as in E, but now for the distance spreads, measuring the variation in the distance between Mic10/Mic19 and Mic60.