M154 media

Primary keratinocytes were isolated as previously described from
human neonatal foreskin obtained from Northwestern University’s Skin
Biology and Disease Resource-based Center (IRB Protocol
#STU00009443).^{33 (link)}Undifferentiated keratinocytes were cultured in M154 media (Thermo Fisher)
supplemented with 0.07mM CaCl₂, human keratinocyte growth
supplement (HKGS), gentamicin, and amphotericin B. To induce differentiation
in 2D cultures, the calcium concentration of the growth medium was raised to
1.2mM CaCl₂. 3D organotypic epidermal raft cultures using primary
keratinocytes were generated as previously described.^{33 (link)} Drug treatments were performed with
200μM Primaquine (Sigma-Aldrich) for 6hr-overnight or 10μM R55
(Sigma-Aldrich) for 48–72hrs. Experiment with pre-differentiation,
keratinocytes were differentiated for 1.5 days before being treated with
200μM Primaquine overnight. During timelapse imaging, keratinocytes
were cultured with FluoroBrite DMEM (Thermo Fisher).

All chemicals were purchased from Sigma Aldrich unless otherwise noted in Supplemental Figure 1. The ASZ and BSZ cell lines were obtained from Professor Robert Holmgren at Northwestern University. FBS, M154 media, DPBS, and NucBlue stain were obtained from ThermoFisher (MT35010CV, M154CF500, 14190250, and R37605, respectively). Trypsin was obtained from Gibco^™ (TrypLE, 12563011) Pen/Strep was obtained from Sigma Aldrich (P0781). All DNA sequences were obtained from IDT with standard desalting and HPLC purification.