antibodies against ager  (R&D Systems)

Journal: bioRxiv

Article Title: Local IFNγ signaling contributes to the regenerative decline of aged alveolar progenitor cells

doi: 10.64898/2026.04.07.716929

Figure Lengend Snippet: Immunoproteasome is required for aged AT2 cell regenerative decline and the establishment of IFN γ+ T cells in tertiary lymphoid structures. A) Schematic figure to show the culture of alveolar organoids from young or aged immunoproteasome knockout (IP KO) and control C57BL/6 (WT) mice in feeder-free media at 7 days of growth, including B) organoid forming efficiency (OFE) and (C) diameter. D) RTqPCR of the IFNγ-inducible MHC-I complex component B2m in isolated lung AT2 cells from young or aged immunoproteasome KO and WT mice. E) Representative immunofluorescence staining of Ki-67 to mark proliferating cells with AT2 cell marker SPC and AT1 cell marker Rage/Ager in corresponding feeder-free organoids from aged mice in A-C. F) The ratio of Ki-67+ proliferating cells in aged immunoproteasome knockout (IP KO) versus WT mice. G) Dysregulated pathways in aged immunoproteasome KO epithelium in comparison to WT based on DEG genes (padj < 0.05, LogFC <0), with bar size representing enrichment score. H) Flow cytometry quantifications of CD8+ and CD4+ (I) T cells as % of all CD45+ immune cells in aged immunoproteasome KO and WT control lungs. J) Zoomed-in multiplex fluorescent images of TLS in aged lungs of immunoproteasome KO and WT controls with K) corresponding absolute quantification of CD8+ T cell and IFNγ+ cell (L) densities. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: We used the following primary antibodies: α-SPC (1:500, AB3786, Millipore), α-RAGE/AGER (1:500, MAB1179-100, R&D System), α-Ki-67 (1:200, 550609, BD Bioscience).

Techniques: Knock-Out, Control, Isolation, Immunofluorescence, Staining, Marker, Comparison, Flow Cytometry, Multiplex Assay, Quantitative Proteomics

Journal: Nature Communications

Article Title: Human iPSC-based Modeling of Pulmonary Fibrosis Reveals p300/CBP Inhibition Suppresses Alveolar Transitional Cell State

doi: 10.1038/s41467-026-68909-z

Figure Lengend Snippet: a Gene expression of ATCS markers in the micro-patterned culture. Each well was treated with either DMSO or p300/CBP inhibitors (10 μM) from days 11 to 14. Data are presented as mean ± SEM (n = 3 biologically independent experiments). One-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01, *** p < 0.001, **** p < 0.0001. b Dot plots displaying the gene expression of iATCs-specific markers in representative cell populations from the micro-patterned culture, corresponding to those shown in Fig. . c Immunostaining of CD54 (ICAM1), lineage markers for ATCS (KRT19), AT1 cells (HT1-56), AT2 cells (NaPi2b), and nuclei (Hoechst) in micro-patterned cultures. Representative images from three biologically independent experiments with similar results are shown. Scale bar: 100 μm. d, e Flow cytometry analysis assessing the CD54 + cell ratio in the micro-patterned culture. Each well was treated with either DMSO or p300/CBP inhibitors (10 μM) from days 11 to 14. Data are presented as mean ± SEM (n = 3 biologically independent experiments). One-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01. f Schematic outline for sorting CD54 + iATCs from day 14 of the micro-patterned culture. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/v51i925 g Gene expression data of ATCS markers in the micro-patterned culture. Data are presented as mean ± SEM ( n = 3 biologically independent experiments). One-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01, *** p < 0.001, **** p < 0.0001. h Schematic outline of the co-culture experiment involving isolated CD54 + iATCs and NHLFs. Created in BioRender. Tsutsui, Y. (2026) https://BioRender.com/nbi6c0b i Gene expression analysis of ATCS markers in isolated CD54 + iATCs. Data are presented as mean ± SEM ( n = 3 biologically independent experiments). One-way ANOVA followed by Tukey’s multiple comparisons test; ** p < 0.01. ns; not significant ( p > 0.05). j Gene expression analysis of fibroblast activation markers in NHLFs cocultured with or without iATCs. Data are presented as mean ± SEM ( n = 3 biologically independent experiments). Unpaired two-tailed Student’s t test: ∗ p < 0.05.

Article Snippet: Primary antibodies used in this study included GFP (1:500, Aves Labs, GFP-1020), SFN (1:200, Abcam, ab77187), act-p300 (1:200, biorbyt, ORB6262), EpCAM (1:200, Santa Cruz Biotechnology, sc-66020), KRT19 (1:200, Merck, MABT913), KRT17 (1:100, Abcam, ab109725), COL1A1 (1:200, Abcam, ab138492), CD54 (1:200, BioLegend, 353102), CD54 (1:200, Atlas, HPA002126), HT1-56 (1:200, Terrace Biotech, TB29AHT1-56), NaPi2b (1:200, kindly provided by Dr. Gerd Ritter (MX35)), AGER (R&D systems, AF1145), H3K27ac (1:200, Cell Signaling technology, 8173), FLAG (1:500, Cell Signaling technology, 14793), alpha smooth muscle Actin (1:100, Abcam ab5694), and Fluorescin (1:1000, Vector Laboratories, FL-1171).

Techniques: Gene Expression, Immunostaining, Flow Cytometry, Co-Culture Assay, Isolation, Activation Assay, Two Tailed Test