Fig. 5: MICU1 and TMBIM5 exhibit reciprocal effects on mitochondrial morphology and submitochondrial localization.

a Morphometric analysis of TMRM labeled mitochondria in WT and T5KO HEK cells expressing control vector (GFP) or MICU1-GFP (M1) imaged with structured illumination microscopy b Morphometric analysis of TMRM labeled mitochondria in WT and M1KO HEK cells expressing control vector (GFP), TMBIM5-GFP (T5), or TMBIM5DM-GFP (T5DM) imaged with structured illumination microscopy. c Analysis of mitochondrial area using TMRM labelling in WT and T5KO HEK cells expressing control vector (GFP) or MICU1-GFP (M1) imaged with structured illumination microscopy. d Analysis of mitochondrial area using TMRM labelling in WT and M1KO HEK cells expressing control vector (GFP), TMBIM5-GFP (T5), or TMBIM5DM-GFP (T5DM) imaged with structured illumination microscopy. e/e’ Analysis of the submitochondrial localization of MICU1-GFP using the IBM association factor with TMRM as reference label in WT and T5KO HEK cells. Representative images of TMRM stained WT and T5KO HEK cells expressing MICU1-GFP (M1) imaged with structured illumination microscopy. f/f´ Analysis of the submitochondrial localization of TMBIM5-GFP and TMBIM5DM-GFP using the IBM association factor with TMRM as reference label in WT and M1KO HEK cells. Representative images of TMRM stained WT and M1KO HEK cells expressing TMBIM5-GFP (T5-GFP) or TMBIM5DM-GFP (T5DM-GFP) imaged with structured illumination microscopy. Data are presented as box and whisker plots with the box representing the interquartile range, spanning from the 25th to the 75th percentile. A horizontal line within the box indicates the median. Whiskers extend from the minimum to the maximum data points. Each data point represents a single cell accumulated from 4 independent experiments. Statistical significance in (a–d and f) was determined using the Kruskal-Wallis test followed by Dunn’s multiple comparisons test and in (e) using unpaired t test, p values are indicated.