Fig. 5: Sec22b controls the levels of phagosomal phospholipids. | Communications Biology

Fig. 5: Sec22b controls the levels of phagosomal phospholipids.

From: Sec22b regulates phagosome maturation by promoting ORP8-mediated lipid exchange at endoplasmic reticulum-phagosome contact sites

Fig. 5

a–c Live spinning-disk confocal microscopy of phagosomal PI(4)P and PI(3)P in shCTR and shSec22b phagocytic MEFs transfected with GFP-2xP4M (green in (a), quantification in (b)) and TagRFP-FYVE(EE1A) (quantification in (c)), exposed to IgG-beads for 30–40 min (n = 8/10 biologically independent samples; 38/36 phagosomes; shCTR/shSec22b). d A similar analysis in cells expressing PI(4,5)P2 probe GFP-C1-PLCdelta-PH (n = 8/6 biologically independent samples; 21/9 phagosomes shCTR/shSec22b). e Quantification of phagosomal PI(4)P in WT phagocytic MEFs expressing GFP-2xP4M and transfected with siCTR or siSec22b (50 nM) (n = 5/5 biologically independent samples; 15/13 phagosomes siCTR/siSec22b; see also Supplementary Fig. S3a). f Quantification of phagosomal PI(4)P in shSec22b phagocytic MEFs transfected with mCh-2xP4M and either GFP-KDEL, shR-EGFP-Sec22b, EGFP-MAPPER or shR-EGFP-Sec22b-P33 (n = 19/14/5/6 biologically independent samples; 100/64/33/24 phagosomes KDEL/Sec22b/ MAPPER/P33; see also Supplementary Fig. S3e). g Quantification of phagosomal PS in shCTR and shSec22b phagocytic MEFs transfected with mRFP-Lact-C2 (n = 5/7 biologically independent samples; 34/52 phagosomes shCTR/shSec22b). White bars = 10 µm. Traces are means ± SEM, and bar graphs are means + SEM.

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