Fig. 2: Lipidomic and immunofluorescence analyses of HeLa cells reveals GM2 accumulation upon loss of progranulin that is restored by PGRN-addback.
From: Deficiency of the frontotemporal dementia gene GRN results in gangliosidosis
a Western blot of full-length progranulin protein levels in GRN+/+, GRN−/−, and GRN−/− + PGRN-addback HeLa cell lines. HeLa cells were gene-edited to contain TMEM192-3xHA. PCNA, proliferating cell nuclear antigen. b Quantification of gangliosides (GM2, GD3, GD1) and c quantification of phospholipids (PC, PE, PS) and neutral lipids (DAG, TAG, SE) isolated from GRN+/+ (green) (n = 7), GRN−/− (orange) (n = 7), and GRN−/− + PGRN-addback (blue) (n = 6) HeLa cell lines. d Representative confocal images of fixed HeLa cells stained with anti-GM2 antibody (magenta), anti-LAMP1 antibody (green) and Hoechst (blue). Scale bar, 50 μm. Bar graphs display number of GM2 puncta per cell and the Pearson’s correlation coefficient between GM2/LAMP1. The numbers of cells used to calculate the GM2 puncta per cell were n = 67, n = 69, n = 63, n = 42 for GRN+/+, GRN−/−, GRN−/− + PGRN-addback, and HEXA−/−, respectively. Box plots display mean ± the minimum and maximum number. One-way ANOVA, followed by multigroup comparison (Dunn’s) test, was performed. *p < 0.05 or ***p < 0.001. PC phosphatidylcholine, PE phosphatidylthanolamine, PS phosphatidylserine, DAG diacylglycerol, TAG triacylglycerol, SE sterol esters.
