Figure 6. SidP does not affect the enzymatic activity of MavQ.

• A
  Direct interactions between SidP and MavQ in the GST pull‐down assay. GST beads coated with GST‐SidP or GST were incubated with His₆‐MavQ. After washing the beads with GST binding buffer, the proteins resolved by SDS–PAGE were detected by Coomassie brilliant blue staining.
• B, C
  SidP does not impair the kinase activity of MavQ in vitro. MavQ was preincubated with increasing amounts of SidP, and the kinase activity of MavQ was measured by the ADP‐Glo assay (B) or the TLC assay (C).
• D
  Co‐expression of SidP with MavQ in HeLa cells does not influence the production of PtdIns3P caused by MavQ. The association of the fluorescence probe GFP‐2xFYVE_Hrs with vesicle‐like structures was used to indicate the distribution of PtdIns3P in cells. At least 100 cells were scored in each sample (n ≥ 100) (left panel). Wort, wortmannin. Expression of MavQ and SidP in the samples was shown in the right panel. EV represents empty vector.
• E
  Deletion of sidP does not impact the association of SidC with the LCV. BMDMs infected with the indicated L. pneumophila stains were stained as described in Fig 5, and the association of SidC with bacterial phagosomes was similarly determined (n ≥ 100) (left panel). Representative images of infected cells harboring bacterial phagosomes were also shown (right panel). Insets represent 3x magnification of regions defined by dash lines. Scale bar, 5 μm.

Data information: In panels A and C, data show one representative of three independent experiments (biological replicates, n = 3). In panel B, data are shown as mean ± SD of triplicates (technical replicates, n = 3) and are representative of three independent experiments (biological replicates, n = 3). In panels D and E, data are shown as mean ± SD of three independent experiments (biological replicates, n = 3). Unpaired two‐tailed Student’s t‐test, p < 0.05 indicates significant difference.

Source data are available online for this figure.