Fig. 5: The expression ratio of anchor and bait protein dictates the function and light responsiveness of protein secretion in LITESEC-act2. | Nature Communications

Fig. 5: The expression ratio of anchor and bait protein dictates the function and light responsiveness of protein secretion in LITESEC-act2.

From: LITESEC-T3SS - Light-controlled protein delivery into eukaryotic cells with high spatial and temporal resolution

Fig. 5

a In vitro secretion assay showing light-dependent export of native T3SS substrates in the LITESEC-act2 strain at different induction levels of anchor expression. For protein assignment and molecular weight, see Fig. 4a. b Quantification of secretion efficiency and light/dark secretion ratio (L/D ratio) for the different expression levels indicated above (as in (a)). N = 3 gel densitometry values from independent experiments for LITESEC-act2 induced with 0.02 or 0.2% arabinose and LITESEC-act3; N = 4 for all other conditions; error bars display standard error of the mean. */**/***p < 0.05/0.01/0.001 in a two-tailed homoscedastic t test (exact values from left to right, 0.95/0.67/0.24/0.012/0.009/0.29/3 × 10−5). c Western blot anti-FLAG of total cellular protein of 2 × 108 bacteria in the LITESEC-act2 strain at the indicated induction levels and conditions. Left side, molecular weight in kDa. See Supplementary Fig. 6 for corresponding Ponceau staining as a loading control. d Correlation between light/dark secretion ratio (L/D ratio) as determined in (b) and anchor expression level. Labels indicate anchor induction levels (arabinose concentrations for LITESEC-act2); the gray dashed line denotes an L/D ratio of 1, indicating light-independent secretion. Source data for panels (b−d) are provided as a Source Data File.

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