Fig. 1: TMBIM5 is upregulated in MCU and MICU1 knockout cells.

a DNA sequencing chromatograms of the genetic locus in wild type (WT) and TMBIM5-knockout (KO) HEK293 cells. The underlined additional adenosine results in a frameshift. b Immunoblot demonstrating loss of TMBIM5 protein expression in KO cells. Actin served as loading control, size is indicated. c Immunoblots demonstrating decreased levels of MCU (upper band, see (e)) and unchanged levels of MICU1, MICU2 and LETM1 in TMBIM5 (T5) KO cells. Actin served as loading control, size is indicated. d Immunoblot demonstrating preserved EMRE protein levels in TMBIM5 KO cells. LETM1 served as loading control for mitochondrial mass and EMRE KO (loaded with higher protein concentration) cells to prove the specificity of the EMRE antibody. Size is indicated. e Immunoblots to demonstrate increased abundance of TMBIM5 in MCU and MICU1 but not MICU2 KO HEK293 cells. Actin served as loading control, size is indicated. Data in c and e are shown as mean ± SD, each data point represents an independent immunoblot. Statistical significance was calculated by the student’s t test in (c) and one-way ANOVA followed by Holm-Šídák’s multiple comparisons test in (e), p values are indicated.