Journal: Frontiers in Immunology

Article Title: MiR-34a deficiency enhances nucleic acid sensing and type I IFN signaling in a mouse model of Alzheimer’s disease

doi: 10.3389/fimmu.2026.1694824

Figure Lengend Snippet: Effects of miR-34a KO on expression levels of homeostatic and DAM genes. Expression levels (Heatmap) of marker genes for three different microglial stages (homeostatic, stage 1 DAM, and stage 2 DAM) are shown as fold changes miR-34a -/- to miR-34a +/+ Tg-SwDI mice, based on bulk RNAseq of the hippocampus (A) . (n=5/group, *Q < 0.05, ##Q < 0.001) Sequence alignment of the mature miR-34a with CSF1R 3’ untranslated region (3’UTR). The seed sequences and target mRNA are highlighted in red (B) . Effects of miR-34a KO on levels of CSF1R in lysates of the cerebral cortex were analyzed by western blotting using CSF1R antibody (C) and bar graph represents the results of densitometric analysis of CSF1R after normalizing with GAPDH levels. Data was analyzed by independent T-test. (n=4/group, ##P < 0.001).

Article Snippet: Membranes were incubated overnight at 4 °C with primary antibodies against IFITM3 (1:1000; Abcam Cat# ab15592, RRID: AB_2122095), CSF1R (1:1000; R and D Systems Cat# AF3818, RRID: AB_884158), and GAPDH (1:5000; Millipore Cat# MAB374, RRID: AB_2107445).

Techniques: Expressing, Marker, RNA sequencing, Sequencing, Western Blot

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Identification of Novel Natural Substrates of Fibroblast Activation Protein-alpha by Differential Degradomics and Proteomics *

doi: 10.1074/mcp.RA118.001046

Figure Lengend Snippet: In vitro examination of candidate substrate CSF-1. A, CSF-1 primary antibody paired with a secondary anti-goat Alexa Fluor 647 stained the N terminus of CSF-1 in the FAP e+ and e− MEFs after permeabilization with 0.05% (w/v) saponin. 23.7% of FAP e− MEFs were immunopositive compared with only 4.5% of the FAP e+ MEFs. Goat IgG was used as a negative control. B, Immunoblotting of CSF-1 in whole cell lysates revealed more cellular CSF-1 present in FAP e− MEFs than FAP e+ MEFs. Densitometry was performed using GAPDH as a loading control. (A, B) Representative of two independent experiments. C, Schematic of the primary structure of CSF-1. The Uniprot-annotated N terminus sequence is shown with the proposed FAP cleavage site indicated by a red arrow after Pro449. This cleavage site identified in TAILS is in the extracellular region, near the transmembrane domain. Identified peptide from TAILS analysis and its corresponding fold change are shown. Synthetic recombinant peptide consists of Ser442 to Arg463, with the proposed FAP cleavage site indicated by a red arrow. D, MALDI-TOF-MS showing no cleavage of the synthetic recombinant CSF-1 peptide after 16 h incubation with rhFAP at 37 °C.

Article Snippet: Primary antibodies were a goat polyclonal to mouse CSF-1 (AF416; R&D Systems) at 0.2 μg/ml and a rabbit polyclonal to LOX-L1 at 0.3 μg/ml (NBP1–82827; Novus Biologicals, Littleton, CO).

Techniques: In Vitro, Staining, Negative Control, Western Blot, Sequencing, Recombinant, Incubation

Journal: Neurobiology of disease

Article Title: Alterations in Alzheimer's disease microglia transcriptome might be involved in bone pathophysiology.

doi: 10.1016/j.nbd.2024.106404

Figure Lengend Snippet: Fig. 3. Elevated levels of CSF1 and Spp1 expression in the 5xFAD mouse brain: Representative image of immunofluorescence staining for co-localization of Iba-1 (green) with (a) CSF1 (red) and (b) Spp1 (red) in the hippocampus sections (Scale bar: 200 μm). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The sections were incubated with primary antibodies (1:200) recognizing CSF1 (Monoclonal Rat IgG2B Clone # 13164 Catalogue Number-MAB4161-SP R&D Systems), SPP1 (Recombinant Monoclonal Rabbit IgG Clone # 2139B Catalohue number-MAB808 R&D Systems) and IBA1 (Ab5076) for 2 h followed by incubation with secondary antibodies (Donkey Anti-Rabbit 650 Catalogue NumberSA5–10041 ThermoFisher Scientific; Donkey Anti-Rat 594 Catalogue Number-A21209 ThermoFisher Scientific; Donkey Anti-Mouse 594 IgG A21203 ThermoFisher Scientific and Anti-Goat 488) for two hours at room temperature.

Techniques: Expressing, Immunofluorescence, Staining