Fig. 1: Interactions of TRiC–PFD with nascent polypeptides.
From: Single-molecule dynamics of the TRiC chaperonin system in vivo
a, Schematic of imaging of TRiC–PFD and ribosomes. b,c, TRiC–ribosome (b) and PFD–ribosome (c) associations. Left, dual-colour imaging. Right, selected time points. Scale bars, 5 µm. d, Duration of chaperone–ribosome interactions (number of cells (n): TRiC, no treatment, n = 26; Puro, n = 26; PFD3 knockdown (KD), n = 26; PFD, no treatment, n = 20; Puro, n = 23). The centre line is the mean and error bars show s.e.m. **P = 0.0086 (one-way Welch’s ANOVA); *P = 0.0314 (Mann–Whitney test). Puro, puromycin. e, Co-movement events per cell (grey) and normalized co-movement events (total interactions divided by number of cells; blue) of interactions as in d. The horizontal line indicates the median, boxes delineate top and bottom quartiles and whiskers extend between 10th and 90th percentiles. ****P < 0.0001 (one-way ANOVA); ****P < 0.0001 (Mann–Whitney test). f, Schematic of dual-colour imaging of TRiC–PFD and translating actin. Actin mRNA is imaged with GFP-tagged MS2 coat protein (GFP–MCP). g, Number of normalized co-movement events (top) and distribution of duration (bottom) for actin–TRiC interactions (number of cells (n): no treatment, n = 38; Puro, n = 40; actin(1–20), n = 42; PFD knockdown, n = 45). The centre line is the mean and error bars show s.e.m. *P = 0.042, **P = 0.006 (one-way Welch’s ANOVA). h, Actin structure (Protein Data Bank (PDB) visualization of AlphaFold predicted structure: AF_AFP60709F1). i, Actin truncations used in experiments. FL, full length. j,k, Normalized frequency of co-movement events (top) and distribution of duration (bottom) of actin truncations with PFD (j) or TRiC (k) (number of cells (n): PFD–actin: actin(1–101), n = 29; actin(1–305), n = 30; actin(1–368), n = 33; actin(FL), n = 38; TRiC–actin: actin(1–101), n = 42; actin(1–168), n = 43; actin(1–305), n = 36; actin(1–368), n = 39; actin(FL), n = 38). The centre line is the mean and error bars show s.e.m. PFD–actin, *P = 0.036, ****P < 0.0001; TRiC–actin, *P = 0.0183, **P = 0.003, ***P = 0.0005 (one-way Welch’s ANOVA).
