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. 2022 Nov 21;41(24):e112677. doi: 10.15252/embj.2022112677

Figure EV5. PI4K3A and PI4K3B do not mediate PtdIns4P recruitment at damaged lysosomes.

Figure EV5

  1. Movie stills from live‐cell imaging experiments of HeLa cells transiently expressing the PtdIns4P probe 2xSidM‐eGFP and the lysosomal marker LAMP1‐mCherry. Cells were incubated with 10‐nM PI4K3A inhibitor GSK‐A1 for approximately 16 min before 250‐μM LLOMe was added.
  2. Movie stills from live‐cell imaging experiments of HeLa cells transiently expressing the PtdIns4P probe 2xSidM‐eGFP and the lysosomal marker LAMP1‐mCherry. Cells were incubated with 25‐nM PI4K3B inhibitor PI4K3B‐IN‐10 for approximately 30 min before 250‐μM LLOMe was added.
  3. Knockdown efficiency of siRNA against PI4K2A or CRISPR‐Cas9‐mediated knockout of PI4K2A (clone 2 and clone 3) as detected by western blot using an anti‐PI4K2A antibody. β‐actin used as a loading control.
  4. Exogenous expression of PI4K2A‐mNG and the PtdIns4P probe SidM‐mCherry in clone 2 and clone 3 of PI4K2A‐KO cells shows recruitment of SidM‐mCherry after incubation with 250‐μM LLOMe.

Source data are available online for this figure.