Extended Data Fig. 5: TFB2M or POLRMT alone could not undergo phase separation.
From: Phase separation drives the self-assembly of mitochondrial nucleoids for transcriptional modulation
a, SDS-PAGE analysis of purified human recombinant POLRMT, TFB2M and TFAM. b, Representative images of POLRMT (2.4 µM) conjugated with Alexa Fluor 647 with or without ND1 DNA (20 nM) under 100 mM NaCl. Scale bar, 10 µm. c, Representative images of TFB2M (10 µM) conjugated with Alexa Fluor 568 with or without ND1 DNA (20 nM) under 100 mM NaCl. Scale bar, 10 µm. d, The incorporation of UTP into the TFAM/DNA droplets. 10 µM UTP-Atto 488 was mixed with 20 µM TFAM and 20 nM D-loop DNA. Scale bar, 5 µm. e, The co-phase separation of UTP into the transcription initiation complex. 10 µM UTP-Atto 488 was mixed with 20 µM TFAM and 20 nM D-loop DNA. Scale bar, 5 µm. f, The phase separation of TFAM truncation mutants, 43-152, 43-222 and 122-246, after adding all the components of transcription initiation complex, including D-loop DNA labeled with YOYO-1, TFB2M and POLRMT conjugated with Alexa Fluor 647. The TFAM (43-246) is set as control. Scale bar, 5 µm. g, The percentage of layered structure of POLRMT of all droplets was counted in left images (f). Data are present with mean ± s.d., and n = 3 independent experiments. Two-tailed unpaired Student’s t-test was used. The p values were noted on the panel. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.
