Journal: bioRxiv

Article Title: Progranulin deficiency induces lipid droplet accumulation in microglia via a STAT3-GPAT3 axis

doi: 10.64898/2026.01.17.700052

Figure Lengend Snippet: a. Volcano plot of differentially expressed genes in Grn -/- BV2 microglial cells. b. GO enrichment analysis of differentially expressed genes (DEGs) from RNA-seq in Grn -/- BV2 microglia. c. Analysis of GPAT3 mRNA levels in Grn -/- BV2 microglial cells (n=3). d. Western blot analysis of GPAT3 and statistical results of gray value quantification in WT and Grn -/- BV2 microglial cells (n=3). e. Analysis of Gpat3 mRNA levels Grn -/- mouse brains at different ages (n=3). f. Representative GPAT3 fluorescence in thalamus of Grn -/- mice at different ages and fluorescence quantification of GPAT3 (n=3). Scale bar, 20 μm. g. Representative BODIPY staining of GPAT3-overexpressing BV2 microglial cells and fluorescence quantification of BODIPY(n=3). Scale bar, 20 μm. h. Representative BODIPY staining of Grn -/- BV2 cells treated with Gpat3 siRNA or scramble siRNA negative control (NC) and fluorescence quantification of BODIPY signals (n=3). Scale bar, 20 μm. i. Representative BODIPY staining of Grn -/- BV2 cells treated with FSG67 or DMSO and fluorescence quantification of BODIPY signals (n=3). Scale bar, 20 μm. j. Western blot analysis of FLAG-tagged PGRN protein in Grn -/- BV2 microglial cells after Grn restoration. k. Analysis of Grn mRNA levels in Grn -/- BV2 microglial cells after Grn restoration (n=6). l-m. Representative GPAT3 fluorescence and BODIPY staining of Grn -/- BV2 microglial cells following PGRN restoration. Scale bar, 20 μm.

Article Snippet: Antibodies used in this experiment are: PGRN (R&D, AF2557, 1:1,000), LAMP1 (Abcam, ab208943, 1:1,000), GPAT3 (Proteintech, 20603-1-AP, 1:2,000), IBA1 (Wako, 019-19741, 1:2,000), GFAP (Cell Signaling Technology, 12389, 1:1,000).

Techniques: RNA Sequencing, Western Blot, Fluorescence, Staining, Negative Control

Journal: bioRxiv

Article Title: Progranulin deficiency induces lipid droplet accumulation in microglia via a STAT3-GPAT3 axis

doi: 10.64898/2026.01.17.700052

Figure Lengend Snippet: a. Venn diagram showing the overlapping of DEGs in four sets of transcriptomic comparison: WT vs. KO2 (Qiantang platform, 1844 DEGs), WT vs. KO2 (Novogene platform, 6770 DEGs), WT vs. KO8 (Qiantang platform, 2127 DEGs), and WT vs. KO8 (Novogene platform, 6042 DEGs). The unique and common DEGs among these comparisons are displayed. Gpat3 is found in the intersection of the four groups. b. Four paired Venn diagrams illustrating the overlap of each individual dataset from with the previously established LD signature genes. The left circle corresponds to the DEG set from , and the right circle corresponds to the list of LD-related marker genes. The overlapping sections highlight shared genes between the DEGs in Grn -/- BV2 microglial cells and the established LD-related genes.

Article Snippet: Antibodies used in this experiment are: PGRN (R&D, AF2557, 1:1,000), LAMP1 (Abcam, ab208943, 1:1,000), GPAT3 (Proteintech, 20603-1-AP, 1:2,000), IBA1 (Wako, 019-19741, 1:2,000), GFAP (Cell Signaling Technology, 12389, 1:1,000).

Techniques: Comparison, Marker

Journal: bioRxiv

Article Title: Progranulin deficiency induces lipid droplet accumulation in microglia via a STAT3-GPAT3 axis

doi: 10.64898/2026.01.17.700052

Figure Lengend Snippet: a. Analysis of Gpat3 mRNA levels in Grn -/- BV2 microglial cells and Grn -/- BV2 cells treated with Gpat3 siRNA1/2/3 (n=3). b. Western blot analysis of GPAT3 in control Grn -/- BV2 cells and Gpat3 siRNA1-treated Grn -/- BV2 cells (n=3). c. Representative PGRN fluorescence a of Grn -/- BV2 microglial cells and Grn -/- Grn OE BV2 cells. Scale bar, 20 μm. d. Analysis of Plin2 and Plin3 mRNA levels in Grn -/- BV2 microglial cells and Grn -/- Grn OE BV2 cells (n=3). e. Analysis of enzymes for triglyceride degradation mRNA levels in Grn -/- BV2 microglial cells and Grn -/- Grn OE BV2 cells (n=3).

Article Snippet: Antibodies used in this experiment are: PGRN (R&D, AF2557, 1:1,000), LAMP1 (Abcam, ab208943, 1:1,000), GPAT3 (Proteintech, 20603-1-AP, 1:2,000), IBA1 (Wako, 019-19741, 1:2,000), GFAP (Cell Signaling Technology, 12389, 1:1,000).

Techniques: Western Blot, Control, Fluorescence

Journal: bioRxiv

Article Title: Progranulin deficiency induces lipid droplet accumulation in microglia via a STAT3-GPAT3 axis

doi: 10.64898/2026.01.17.700052

Figure Lengend Snippet: a. Gene se enrichment analysis (GSEA) of IL-6 signaling in Grn -/- BV2 microglial cells. b. Analysis of IL-6 mRNA levels in Grn -/- BV2 microglial cells and Grn -restored Grn -/- BV 2 microglial cells (n=6). c. Analysis of IL-6 mRNA levels in Grn -/- mouse brains at different ages (n=3). d. Western blot analysis of p-STAT3(tyr705) and statistical results of gray value quantification. in Grn -/- BV2 microglial cells and Grn -restored Grn -/- BV2 microglial cells (n=4). e. Western blot analysis of p-STAT3(tyr705) and p-STAT3(ser727) and statistical results of gray value quantification 9-month-old Grn -/- mice (n=4). f. Western blot analysis of p-STAT3(tyr705) p-STAT3(ser727) and statistical results of gray value quantification in Grn -/- BV2 microglial cells and Grn -/- BV2 microglial cells treated with Stattic (n=3). g. Analysis of GPAT3 mRNA levels in Grn -/- BV2 microglial cells and Grn -/- BV2 microglial cells treated with Stattic (n=6). h. Western blot analysis of GPAT3 and statistical results of gray value quantification in Grn -/- BV2 microglial cells and Grn -/- BV2 microglial cells treated with Stattic (n=3). i. Representative BODIPY staining of Grn -/- BV2 microglial cells and Grn -/- BV2 microglial cells treated with Stattic and fluorescence quantification of BODIPY (n=3). Scale bar, 20 μm.

Article Snippet: Antibodies used in this experiment are: PGRN (R&D, AF2557, 1:1,000), LAMP1 (Abcam, ab208943, 1:1,000), GPAT3 (Proteintech, 20603-1-AP, 1:2,000), IBA1 (Wako, 019-19741, 1:2,000), GFAP (Cell Signaling Technology, 12389, 1:1,000).

Techniques: Western Blot, Staining, Fluorescence

Journal: bioRxiv

Article Title: Progranulin deficiency induces lipid droplet accumulation in microglia via a STAT3-GPAT3 axis

doi: 10.64898/2026.01.17.700052

Figure Lengend Snippet: a. Cell viability assay of mouse neuronal N2a cells treated with conditioned media from either BV 2 microglia cells or Grn -/- BV2 microglial cells (n = 5). b. Quantitative analysis of intracellular lactate dehydrogenase (LDH) levels in N2a cells treated as in (A) (n = 5). c. Reactive oxygen species (ROS) levels in N2a cells treated as in (A) (n = 5). d. Cell viability assay of mouse neuronal N2a cells treated with conditioned media from either Grn -/- microglial cells or Grn -rescued Grn -/- microglia (n = 5). e. Intracellular reactive oxygen species (ROS) levels in mouse neurons treated as in (D) (n = 5). f. N2a cell viability after treatment with conditioned media from either Grn -/- microglial cells or siRNA- Gpat3 treated Grn -/- microglial cells (n = 4). g. N2a cell viability after treatment with conditioned media from either Grn -/- microglial cells or FSG67-treated Grn -/- microglial cells (n = 3).

Article Snippet: Antibodies used in this experiment are: PGRN (R&D, AF2557, 1:1,000), LAMP1 (Abcam, ab208943, 1:1,000), GPAT3 (Proteintech, 20603-1-AP, 1:2,000), IBA1 (Wako, 019-19741, 1:2,000), GFAP (Cell Signaling Technology, 12389, 1:1,000).

Techniques: Viability Assay

Journal: bioRxiv

Article Title: Progranulin deficiency induces lipid droplet accumulation in microglia via a STAT3-GPAT3 axis

doi: 10.64898/2026.01.17.700052

Figure Lengend Snippet: a. Representative GPAT3 fluorescence in thalamus of Grn -/- mice injected with AAV-CTRL or AAV- Grn and fluorescence quantification of GPAT3 signals (n = 3). Scale bar, 20 μm. b. Representative LipidSpot staining in thalamus of Grn -/- mice injected with AAV-CTRL or AAV- Grn and fluorescence quantification of LipidSpot (n = 3). Scale bar, 20 μm. c. Representative IBA1 fluorescence in thalamus of Grn -/- mice injected with AAV-CTRL or AAV- Grn and fluorescence quantification of IBA1 signals (n = 3). Scale bar, 20 μm. d. Representative GFAP fluorescence in thalamus of Grn -/- mice injected with AAV-CTRL or AAV- Grn and fluorescence quantification of GFAP signals (n = 3). Scale bar, 20 μm. e. Representative BODIPY and IBA1 fluorescence staining in thalamus of Grn -/- mice treated with FSG67 or vehicle and fluorescence quantification of BODIPY in IBA + cells (n = 3). Scale bar, 20 μm. f. Recognition working memory assessed by the novel object recognition test in WT and Grn -/- mice treated with FSG67 or vehicle (n = 5 for each group). g. Anxiety behavior assessed by the light-dark transition test in WT and Grn -/- mice treated with FSG67 or vehicle (n = 5 for each group). h. Anxiety behavior assessed by the open field test in WT and Grn -/- mice treated with FSG67 or vehicle (n = 5 for each group). i. Representative open-field trajectory heatmaps for the groups in (SuperMaze software). j. Recognition working memory assessed by the novel object recognition test in Grn -/- mice injected with AAV-CTRL or AAV- Grn (n = 7 for each group). k. Anxiety behavior assessed by the light-dark transition test in Grn -/- mice injected with AAV-CTRL or AAV- Grn (n = 7 for each group). l. Anxiety behavior assessed by the open field test in Grn -/- mice injected with AAV-CTRL or AAV- Grn (n = 7 for each group). m. Representative open-field trajectory heatmaps for the groups in (AnyMaze software).

Article Snippet: Antibodies used in this experiment are: PGRN (R&D, AF2557, 1:1,000), LAMP1 (Abcam, ab208943, 1:1,000), GPAT3 (Proteintech, 20603-1-AP, 1:2,000), IBA1 (Wako, 019-19741, 1:2,000), GFAP (Cell Signaling Technology, 12389, 1:1,000).

Techniques: Fluorescence, Injection, Staining, Software