Journal: Npj Dementia

Article Title: Progranulin deficiency does not exacerbate TDP-43 pathology in TDP-43 transgenic mouse models

doi: 10.1038/s44400-025-00020-4

Figure Lengend Snippet: A Immunostaining of TDP-43 and NeuN in brain sections from 16-month-old mice WT, TDP-43 Q331K/Q331K (Q331K), Grn −/− , TDP-43 Q331K/Q331K Grn −/− (Q331K Grn −/− ) mice. Representative images from the cortex were shown. Scale bar, 10 µm. B Analysis of TDP-43, phosphorylated TDP-43 (pS409/410), and PGRN levels in cortical lysates from 16-month-old mice of the indicated genotypes. C TDP-43 levels in RIPA- and urea-soluble fractions were quantified. Data are presented as mean ± SEM from 3 mice per group ( n = 3). One-way ANOVA tests with Bonferroni’s multiple comparisons. D PGRN levels in RIPA soluble fractions were quantified and normalized to GAPDH. Data are presented as mean ± SEM from 3 mice per group ( n = 3). E Total RNAs were extracted from the cortex of 10-month-old WT, TDP-43 Q331K/Q331K , Grn −/− , TDP-43 Q331K/Q331K Grn −/− male mice, and the RT-qPCR was performed to analyze the splicing changes in Sort1 exon 17b (left) and Mapt exons 2 and 3 (right). The relative mRNA levels of transcripts including or excluding exons 2 and 3 represent the inclusion of Mapt exons 2 and 3. Data are presented as mean ± SEM ( n = 4 mice per genotype). p -values were determined using one-way ANOVA tests with Bonferroni’s multiple comparisons. F Expression levels of Tardbp in WT and Q331K mice. Total RNAs were extracted from the cortex of 10-month-old WT and Q331K male mice, and the RNA-seq was performed to analyze gene expression changes. Normalized read counts are shown. Data are presented as mean ± SEM ( n = 5-6 mice per genotype). * p < 0.05, unpaired two-tailed Student's t-test.

Article Snippet: The following antibodies were used in this study: rabbit anti-IBA-1 (Wako, 01919741), goat anti-AIF-1/Iba1 (Novus Biologicals, NB100-1028), rat anti-CD68 (Bio-Rad, MCA1957), mouse anti-GFAP (Cell signaling, 3670S), rabbit anti-TDP43 (Proteintech Group, 12892-1-AP (C-terminal) and 10782-2-AP (N-terminal)), rabbit anti-phospho-TDP-43 (Ser409/410) (Proteintech group, 80007-1-RR), mouse anti-PLP (Millipore, MAB388), mouse anti-MBP (Millipore, SMI-99), rabbit anti-MAG (Proteintech Group, 14386-1-AP), sheep anti-mouse PGRN (R&D systems, AF2557), and mouse anti-GAPDH (Proteintech Group, 60004-1-Ig).

Techniques: Immunostaining, Quantitative RT-PCR, Expressing, RNA Sequencing, Gene Expression, Two Tailed Test

Journal: Npj Dementia

Article Title: Progranulin deficiency does not exacerbate TDP-43 pathology in TDP-43 transgenic mouse models

doi: 10.1038/s44400-025-00020-4

Figure Lengend Snippet: A Immunostaining of TDP-43 and NeuN using rabbit anti-TDP-43 CTD antibodies and mouse anti-NeuN antibodies, respectively (left panel) or immunostaining of human TDP-43 (right panel) using mouse anti-human TDP-43 in brain sections from 21-day-old WT, hTDP-43 Tg/Tg (Tg/Tg), Grn −/− , and hTDP-43 Tg/Tg Grn −/− mice. Representative images from the cortex were shown. Scale bar, 10 µm. B Analysis of TDP-43 and phosphorylated TDP-43 (pS409/410) and PGRN levels in brain lysates from 21-day-old mice. Antibodies recognizing the C-terminal or N-terminal domain of TDP-43 were used to detect total TDP-43 levels. C TDP-43 and pTDP-43 levels in RIPA- and urea-soluble fractions were quantified. Data are presented as mean ± SEM from 3 mice per group ( n = 3). One-way ANOVA tests with Bonferroni’s multiple comparisons. ** p < 0.01. D PGRN levels in RIPA soluble fractions were quantified and normalized to GAPDH. Data are presented as mean ± SEM from 3 mice per group ( n = 3).

Article Snippet: The following antibodies were used in this study: rabbit anti-IBA-1 (Wako, 01919741), goat anti-AIF-1/Iba1 (Novus Biologicals, NB100-1028), rat anti-CD68 (Bio-Rad, MCA1957), mouse anti-GFAP (Cell signaling, 3670S), rabbit anti-TDP43 (Proteintech Group, 12892-1-AP (C-terminal) and 10782-2-AP (N-terminal)), rabbit anti-phospho-TDP-43 (Ser409/410) (Proteintech group, 80007-1-RR), mouse anti-PLP (Millipore, MAB388), mouse anti-MBP (Millipore, SMI-99), rabbit anti-MAG (Proteintech Group, 14386-1-AP), sheep anti-mouse PGRN (R&D systems, AF2557), and mouse anti-GAPDH (Proteintech Group, 60004-1-Ig).

Techniques: Immunostaining

Journal: eNeuro

Article Title: Effects of Exercise on Progranulin Levels and Gliosis in Progranulin-Insufficient Mice 1 , 2 , 3

doi: 10.1523/ENEURO.0061-14.2015

Figure Lengend Snippet: Exercise (3-4 weeks) produces a small increase in hippocampal progranulin in young adult wild-type mice. Two- to 3-month-old wild-type mice were randomized to sedentary or exercise groups for three to four weeks ( A ). B , Exercise produced a robust increase in BDNF (**** p < 0.0001). C , Representative BDNF and α-tubulin blots. D , Exercise did not significantly increase frontal cortex progranulin protein. E , Exercise produced a small but statistically significant increase in hippocampal progranulin (* p = 0.038). F , Exercise did not affect plasma progranulin. n = 8–16 mice per group. Values in B – F are expressed relative to the sedentary group.

Article Snippet: After electrophoresis, transfer, and blocking as described above, the samples were incubated overnight in a sheep polyclonal anti-progranulin antibody (1:5000, R&D Systems AF2557).

Techniques: Produced, Clinical Proteomics

Journal: eNeuro

Article Title: Effects of Exercise on Progranulin Levels and Gliosis in Progranulin-Insufficient Mice 1 , 2 , 3

doi: 10.1523/ENEURO.0061-14.2015

Figure Lengend Snippet: Exercise (4 weeks) does not increase progranulin protein levels in multiple brain regions of Grn +/− mice. Four- to 8-month-old Grn +/− mice were randomized to sedentary or exercise groups for 4 weeks ( A ). Exercise did not increase progranulin protein levels in frontal cortex ( B ), thalamus ( C ), or hippocampus ( D ), despite producing the expected increase in the number of doublecortin-positive neurons in the dentate gyrus ( E , ** p < 0.01). Values in B – D are expressed relative to sedentary Grn +/+ mice, with sedentary Grn +/− mice set at 0.5 to maintain a consistent scale with Figure 3. n = 11–12 per group.

Article Snippet: After electrophoresis, transfer, and blocking as described above, the samples were incubated overnight in a sheep polyclonal anti-progranulin antibody (1:5000, R&D Systems AF2557).

Techniques:

Journal: eNeuro

Article Title: Effects of Exercise on Progranulin Levels and Gliosis in Progranulin-Insufficient Mice 1 , 2 , 3

doi: 10.1523/ENEURO.0061-14.2015

Figure Lengend Snippet: Measures of progranulin levels

Article Snippet: After electrophoresis, transfer, and blocking as described above, the samples were incubated overnight in a sheep polyclonal anti-progranulin antibody (1:5000, R&D Systems AF2557).

Techniques: Clinical Proteomics

Journal: eNeuro

Article Title: Effects of Exercise on Progranulin Levels and Gliosis in Progranulin-Insufficient Mice 1 , 2 , 3

doi: 10.1523/ENEURO.0061-14.2015

Figure Lengend Snippet: Group housing does not potentiate the effects of exercise on progranulin. Six weeks of wheel running ( A ) failed to increase frontal cortex progranulin mRNA ( B ) or hippocampal progranulin protein ( C ) in either solo- or group-housed wild-type mice aged 3–6 months. D-F , Hippocampal BDNF was increased by exercise ( D ; ANOVA effect of exercise, p = 0.041). F , Representative BDNF and α-tubulin blots for each group. n = 8–14 mice per group. Values in B – E are expressed relative to the solo-housed sedentary group.

Article Snippet: After electrophoresis, transfer, and blocking as described above, the samples were incubated overnight in a sheep polyclonal anti-progranulin antibody (1:5000, R&D Systems AF2557).

Techniques:

Journal: The American journal of pathology

Article Title: Accelerated lipofuscinosis and ubiquitination in granulin knockout mice suggest a role for progranulin in successful aging.

doi: 10.2353/ajpath.2010.090915

Figure Lengend Snippet: Figure 1. PGRN immunoreactivity is reflective of genotypes and increases in older GRN/ and GRN/ mice. As shown in 12-month-old mice, GRN/ mice (A) have higher PGRN immunoreactivity than GRN/ mice (B), whereas GRN/ mice (C) have no detectable PGRN expression. Higher magnification insets correspond to the areas identified by black squares. D: In a subset of GRN/ (green) and GRN/ (orange) mice, PGRN immuno- reactivity was quantified by image analysis. Consistent with genotypes, GRN/ mice had significantly reduced PGRN levels compared with GRN/

Article Snippet: Membranes were blocked in 5% milk in Tris-buffered saline-0.1% Triton X-100 and incubated overnight in anti-mouse PGRN (1:500; R&D Systems, Minneapolis, MN) or anti-glyceraldehyde-3-phosphate dehydrogenase (1:10,000; BioSource International, Camarillo, CA) at 4°C.

Techniques: Expressing

Journal: The American journal of pathology

Article Title: Accelerated lipofuscinosis and ubiquitination in granulin knockout mice suggest a role for progranulin in successful aging.

doi: 10.2353/ajpath.2010.090915

Figure Lengend Snippet: Figure 7. Hypothetical model of neurodegeneration based on the chrono- logical appearance of pathology in GRN/ mice. 1 month: young neurons are free of any pathology and microglia are in a ramified or resting state; seven months: through an unknown mechanism, the lack of PGRN results in intraneuronal accumulation of lipofuscin pigment in aging neurons resulting in cellular stress which is detected by nearby microglia; 12 months: neuronal cytoplasm is full of lipofuscin pigment which adversely affects cellular func- tion and long-term neuronal survival, signaling the activation of microglia; 23 months: slow and progressive loss of neurons, resulting in expulsion of neuronal lipofuscin into extracellular space, most of which is phagocytosed by activated microglia with some lipofuscin accumulating in the extracellular space. The lack of PGRN may also affect the normal function of activated microglia resulting in accumulation of endogenous microglial lipofuscin. A severe, sustained, and/or an inappropriate microglia (inflammatory) re- sponse (attributable to absence of microglial PGRN) may further enhance neuronal loss.

Article Snippet: Membranes were blocked in 5% milk in Tris-buffered saline-0.1% Triton X-100 and incubated overnight in anti-mouse PGRN (1:500; R&D Systems, Minneapolis, MN) or anti-glyceraldehyde-3-phosphate dehydrogenase (1:10,000; BioSource International, Camarillo, CA) at 4°C.

Techniques: Activation Assay

Journal: Stem Cells Translational Medicine

Article Title: Progranulin, a Major Secreted Protein of Mouse Adipose-Derived Stem Cells, Inhibits Light-Induced Retinal Degeneration

doi: 10.5966/sctm.2013-0020

Figure Lengend Snippet: Progranulin suppressed H2O2- or light-induced cultured photoreceptor cell death. (A, B): Quantitative determination of progranulin in ASC-CM by Western blot. (A): Representative immunoblots showing progranulin protein levels in ASC-CM and recombinant mouse progranulin. The recombinant progranulin was expressed in a mouse myeloma cell line, and post-translational modifications such as glycosylation may contribute to various molecular weights. (B): Standard curve was generated from the density of mouse recombinant progranulin. (C, E): Representative fluorescence microscopy images showing nuclear staining for Hoechst 33342 and PI after 27 hours of H2O2 (0.3 mM) treatment (C) or 24 hours of light exposure (E). (D, F): The number of cells exhibiting PI fluorescence was counted, and positive cells were expressed as the percentage of PI- to Hoechst 33342-positive cells. The number of PI-positive cells increased after H2O2 treatment or light exposure. Progranulin significantly reduced cell death in a concentration-dependent manner. Scale bars = 50 µm. Data are shown as mean ± SEM (n = 6 or n = 9). *, p < .05 versus vehicle; **, p < .01; ##, p < .01 versus control. Abbreviations: ASC-CM, adipose-derived stem cell-conditioned medium; Con, control; H2O2, hydrogen peroxide; PI, propidium iodide; Veh, vehicle.

Article Snippet: For immunoblotting, the following primary antibodies were used: monoclonal anti-mouse progranulin antibody (R&D Systems), anti-β-actin, (Sigma-Aldrich), anti-ERK1/2, anti-phosphorylated ERK1/2, anti-HGF receptor (MET), anti-phosphorylated MET (Cell Signaling Technology, Danvers, MA, http://www.cellsignal.com ), anti-cAMP response element binding protein (Cell Signaling Technology), and anti-phosphorylated CREB (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, http://www.scbt.com ).

Techniques: Cell Culture, Western Blot, Recombinant, Glycoproteomics, Generated, Fluorescence, Microscopy, Staining, Concentration Assay, Control, Derivative Assay

Journal: Stem Cells Translational Medicine

Article Title: Progranulin, a Major Secreted Protein of Mouse Adipose-Derived Stem Cells, Inhibits Light-Induced Retinal Degeneration

doi: 10.5966/sctm.2013-0020

Figure Lengend Snippet: Progranulin reduced retinal damage induced by exposure to light in mice. (A): Representative photographs of hematoxylin and eosin staining are as follows: nontreated group (Aa), light exposure (8,000 lx) plus vehicle-treated group (Ab), and light exposure plus progranulin-treated (200 ng per eye) group (Ac) at 5 days after light exposure. (B): ONL thickness was measured at 5 days after light exposure. The ONL was measured at 240-μm intervals from the optic disc. Scale bar = 25 µm. Data are shown as mean ± SEM (n = 7 or n = 8). *, p < .05 versus the light exposure plus vehicle-treated group (vehicle). Abbreviation: ONL, outer nuclear layer.

Article Snippet: For immunoblotting, the following primary antibodies were used: monoclonal anti-mouse progranulin antibody (R&D Systems), anti-β-actin, (Sigma-Aldrich), anti-ERK1/2, anti-phosphorylated ERK1/2, anti-HGF receptor (MET), anti-phosphorylated MET (Cell Signaling Technology, Danvers, MA, http://www.cellsignal.com ), anti-cAMP response element binding protein (Cell Signaling Technology), and anti-phosphorylated CREB (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, http://www.scbt.com ).

Techniques: Staining

Journal: Stem Cells Translational Medicine

Article Title: Progranulin, a Major Secreted Protein of Mouse Adipose-Derived Stem Cells, Inhibits Light-Induced Retinal Degeneration

doi: 10.5966/sctm.2013-0020

Figure Lengend Snippet: Progranulin exerted a photoreceptor protective effect through the protein kinase C pathway. Representative band images show immunoreactivity against pERK, total ERK, pCREB, and total CREB (A) and phosphorylated HGFR and total HGFR (D) after progranulin treatment (500 ng/ml) at 5, 10, 30, and 60 minutes. (B, E): Quantitative analysis of band densities. Data are shown as mean ± SEM (n = 3). (C): Progranulin (500 ng/ml), U0126 (1,000 nM), H-87 (500 nM), and Gö 6976 (500 nM) were added to 661W cells, and the cells were irradiated with white light (2,500 lx) for 24 hours. Cell death rate was calculated as described in Materials and Methods. Data are shown as mean ± SEM (n = 6). *, p < .05 versus vehicle; **, p < .01. Abbreviations: CREB, cAMP response element binding protein; HGFR, hepatocyte growth factor receptor; n.s., not significant; P, progranulin; pCREB, phosphorylated CREB; pERK, phosphorylated ERK; V, vehicle.

Article Snippet: For immunoblotting, the following primary antibodies were used: monoclonal anti-mouse progranulin antibody (R&D Systems), anti-β-actin, (Sigma-Aldrich), anti-ERK1/2, anti-phosphorylated ERK1/2, anti-HGF receptor (MET), anti-phosphorylated MET (Cell Signaling Technology, Danvers, MA, http://www.cellsignal.com ), anti-cAMP response element binding protein (Cell Signaling Technology), and anti-phosphorylated CREB (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, http://www.scbt.com ).

Techniques: Irradiation, Binding Assay