Table 1. Intra-Golgi transport kinetics of secretory RUSH reporters in Golgi ministacks.
See the legend of Figure 2 for details. #1–4 indicate independent replicates. All data were acquired from HeLa cells except for those labeled with ‘293T,’ which were acquired using 293T cells. Except for TNFα-SBP-GFP and SBP-GFP, which use Ii-streptavidin as the ER hook, all RUSH reporters employ signal sequence fused streptavidin-KDEL as their ER hook. Superscripts 1 and 2 indicate data were re-analyzed from previous publications, (Tie et al., 2016) and (Sun et al., 2020), respectively. WT, wild type. tintra (intra-Golgi transport time), y0, and adj. R2 was calculated by fitting measured LQ vs. time kinetics data to Equation 1. dLQ/dt (at LQ = 0.40) was calculated by Equation 3, and converted to nm/min by multiplying 274 nm per LQ unit. SD, standard deviation.
| RUSH reporter | tintra (min) Mean ±SD | tintra (min) | y 0 | Adj. R2 | dLQ/dt (at LQ = 0.40) | dLQ/dt (at LQ = 0.40) (nm/min) |
|---|---|---|---|---|---|---|
| SBP-GFP-CD59 #1 (293T) | 5±4 | 0.4 | 0.66 | 0.94 | 0.244 | 66.7 |
| SBP-GFP-CD59 #2 (293T) | 7.3 | 0.63 | 0.92 | 0.022 | 6.0 | |
| SBP-GFP-CD59 #3 (293T) | 6.8 | 0.79 | 0.85 | 0.040 | 10.9 | |
| TNFα-SBP-GFP1 | 6.4 | 6.4 | 1.09 | 0.94 | 0.075 | 20.5 |
| TfR-SBP-GFP | 7 | 7.0 | 1.16 | 0.99 | 0.075 | 20.6 |
| SBP-GFP-CD8a-furin-Y+AC #11 | 7±1 | 4.9 | 0.83 | 0.99 | 0.061 | 16.7 |
| SBP-GFP-CD8a-furin-Y+AC #21 | 7.1 | 1.00 | 0.99 | 0.059 | 16.0 | |
| SBP-GFP-CD8a-furin-Y+AC #31 | 7.7 | 1.04 | 0.92 | 0.058 | 15.8 | |
| SBP-GFP-Tac #12 | 7±2 | 6.0 | 0.86 | 0.98 | 0.053 | 14.6 |
| SBP-GFP-Tac #2 | 8.2 | 0.91 | 0.98 | 0.043 | 11.8 | |
| SBP-GFP-E-cadherin #11 | 8±1 | 7.6 | 0.98 | 0.99 | 0.053 | 14.5 |
| SBP-GFP-E-cadherin #21 | 8.7 | 1.15 | 0.96 | 0.060 | 16.5 | |
| SBP-GFP-E-cadherin #31 | 6.5 | 1.11 | 0.98 | 0.075 | 20.6 | |
| SBP-GFP-collagenX #1 | 8±1 | 9.0 | 1.00 | 0.94 | 0.046 | 12.7 |
| SBP-GFP-collagenX #2 | 9.5 | 0.84 | 0.97 | 0.032 | 8.8 | |
| SBP-GFP-collagenX #3 | 6.9 | 0.91 | 0.97 | 0.051 | 14.0 | |
| SBP-GFP | 10.6 | 10.6 | 0.74 | 0.98 | 0.022 | 6.1 |
| SBP-GFP-CD8a-furin-YA #11 | 11±5 | 5.9 | 1.49 | 0.98 | 0.13 | 35.0 |
| SBP-GFP-CD8a-furin-YA #21 | 11.1 | 1.51 | 0.93 | 0.069 | 19.0 | |
| SBP-GFP-CD8a-furin-YA #31 | 16.3 | 1.48 | 0.95 | 0.046 | 12.6 | |
| SBP-GFP-CD591 | 13.0 | 13.0 | 1.03 | 0.97 | 0.034 | 9.2 |
| SBP-GFP-Tac-TC #1 (293T) | 14.2±0.6 | 14.7 | 0.95 | 0.97 | 0.026 | 7.1 |
| SBP-GFP-Tac-TC #2 (293T) | 14.3 | 1.00 | 0.98 | 0.029 | 8.0 | |
| SBP-GFP-Tac-TC #3 (293T) | 13.6 | 0.98 | 0.89 | 0.030 | 8.1 | |
| SBP-GFP-Tac-TC #12 | 17±3 | 15.5 | 1.05 | 0.97 | 0.029 | 8.0 |
| SBP-GFP-Tac-TC #2 | 19.1 | 1.16 | 0.94 | 0.028 | 7.6 | |
| SBP-GFP-CD8a-furin-WT #11 | 18±5 | 23.6 | 1.63 | 0.96 | 0.036 | 9.9 |
| SBP-GFP-CD8a-furin-WT #21 | 18.7 | 1.58 | 0.97 | 0.044 | 12.0 | |
| SBP-GFP-CD8a-furin-WT #31 | 13.1 | 1.55 | 0.97 | 0.061 | 16.7 | |
| SBP-GFP-CD8a-furin-AC #11 | 21±5 | 17.9 | 1.73 | 0.99 | 0.052 | 14.1 |
| SBP-GFP-CD8a-furin-AC #21 | 26.9 | 1.50 | 1.00 | 0.028 | 7.8 | |
| SBP-GFP-CD8a-furin-AC #31 | 23.2 | 1.58 | 0.96 | 0.035 | 9.7 | |
| SBP-GFP-CD8a-furin-AC #41 | 16.8 | 1.62 | 0.99 | 0.050 | 13.8 |