olfm4 antibody  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: Fasting primes small intestinal regeneration after damage via a microbiome–metabolite-chromatin axis

doi: 10.64898/2026.03.06.710208

Figure Lengend Snippet: a. Schema of experimental design. C57BL/6J mice were administered antibiotics (Ampicillin 0.5 g/L and Enrofloxacin 0.25 g/L) in drinking water for 7 days followed by three days of antibiotic clearance. AKK (1×10 8 CFU) or vehicle (PBS) were introduced by gavage on day -7 and day -4. On day -1, mice were fed or fasted for 24 h and then exposed to total abdominal radiation (11.5 Gy; D0). Mice were then housed singly with food and on day 5 post radiation a cohort of mice was sacrificed, and ileums isolated for analysis. Remaining mice were monitored for survival until day 30 post radiation. Ileums were also isolated from mice on D-10 and D0 for 16S rRNA sequencing. b. Relative abundances of bacteria at the genus level in ileum samples collected on D0. Each column represents an individual mouse. c. Survival of mice following TA-XRT was monitored daily for 30 days. P= 0.0023 for FED (- AKK ) vs FAST (+ AKK ), P= 0.8488 for FED (- AKK ) vs FED (+ AKK ), P= 0.2698 for FED (- AKK ) vs FAST (- AKK ) (using Logrank (Mantel-Cox) test) d. Representative images of H & E-stained ileum collected on Day 5. Scale bars, 200 µm. Magnification, 20x. Representative images shown from 4 mice. e. Number of crypts per 2 mm of SI (ileum) and their depth were quantitated for each day 5 sample. The average number of crypts per mm of ileum length and the average crypt depth were plotted (n=10 fields per mouse). ns nonsignificant; *P<0.05; ****P<0.0001 by one-way ANOVA. Error bars are ± SEM. f. Representative images of ileum stained for Olfm4 (day 5 samples). Scale bars, 200 µm. Magnification, 20x. Representative images shown for 4 mice. g. Percentage of crypt epithelial cells staining positive for Olfm4 is shown. ns, nonsignificant; ****P<0.0001 by one-way ANOVA. Error bars are ± SEM. 10 fields per mouse form 4 mice.

Article Snippet: Tissues were blocked with Protein Block (Dako # X0909) and/or normal serum (Vector ImmPRESS # MP-7401), followed by overnight incubation with 1:200 dilution of Olfm4 antibody (CST # 39141S) at 4°C.

Techniques: Isolation, Sequencing, Bacteria, Staining

Journal: bioRxiv

Article Title: Fasting primes small intestinal regeneration after damage via a microbiome–metabolite-chromatin axis

doi: 10.64898/2026.03.06.710208

Figure Lengend Snippet: a. Schema of experimental design. C57BL/6J mice were administered vehicle or antibiotics (Tetracycline 3 g/L and 10% sucrose) in drinking water for 21 days followed by three days of antibiotic clearance. AKK (1×10 8 CFU) or vehicle (PBS) were introduced by gavage on day -7 and day -4. On day -1, mice were fed or fasted for 24 h and then exposed to total abdominal radiation (11.5 Gy; D0). Mice were then housed singly with food and on day 5 a cohort of mice was sacrificed, and ileums isolated for analysis. Remaining mice were monitored for survival until day 30 post radiation. Ileums were also isolated from mice on D-10 and D0 for 16S rRNA sequencing. b. Relative abundances of bacteria at the genus level in ileum samples collected at D0. Each column represents an individual mouse. c. Survival of mice following TA-XRT was monitored daily for 30 days. P= 0.0038 for FED (- AKK ) vs FAST (+ AKK ), P= 0.1036 for FED (- AKK ) vs FED (+ AKK ), P= 0.4313 for FED (- AKK ) vs FAST (- AKK ) (using Logrank (Mantel-Cox) test) d. Representative images of H & E-stained ileum collected on Day 5. Scale bars, 200 µm. Magnification, 20x. Representative images shown from 4 mice. e. Number of crypts per 2 mm of SI (ileum) and their depth was quantitated for each day 5 sample. The average number of crypts per mm of ileum length and the average crypt depth was plotted (n=10 fields per mouse). ns, nonsignificant; ***P<0.0003; ****P<0.0001 by one-way ANOVA. Error bars are ± SEM. f. Representative images of ileum stained for Olfm4 (day 5 samples). Scale bars, 200 µm. Magnification, 20x. Representative images shown for 4 mice. g. Percentage of crypt epithelial cells staining positive for Olfm4 is shown. ns, nonsignificant, ****P<0.0001 by one-way ANOVA. Error bars are ± SEM. 10 fields per mouse form 4 mice.

Article Snippet: Tissues were blocked with Protein Block (Dako # X0909) and/or normal serum (Vector ImmPRESS # MP-7401), followed by overnight incubation with 1:200 dilution of Olfm4 antibody (CST # 39141S) at 4°C.

Techniques: Isolation, Sequencing, Bacteria, Staining

Journal: bioRxiv

Article Title: Fasting primes small intestinal regeneration after damage via a microbiome–metabolite-chromatin axis

doi: 10.64898/2026.03.06.710208

Figure Lengend Snippet: a. Experimental design. Mice were fed ad libitum or fasted for 24 h, followed by total abdominal irradiation (11.5 Gy). Small intestinal crypt cells were collected at 0 h, 24 hpi, 48 hpi, or 72 hpi (post-irradiation) and subjected to single-cell ATAC sequencing (scATAC-seq). b. UMAP projection of aggregated scATAC-seq data colored by sample, showing condition-specific shifts in chromatin accessibility. c. UMAP colored by cluster identity, identifying major intestinal epithelial populations. d. Bubble plot showing chromatin accessibility of stem and revival marker loci (Clu, Olfm4) across conditions. Cluster 6 corresponds to the Clu+Olfm4+ persister stem cell population. e. Bubble plot showing chromatin accessibility (log2FC) of stem cell, revival, and injury-associated gene loci in Cluster 3 (C3) and Cluster 6 (C6). f. Percentage of Clu+Olfm4+ cells (Cluster 6) across all samples, demonstrating increased representation during fasting. g. Integration of CUT&Tag H3K27ac data with RNA-seq for the activated genes in Fed (-Tetra), Fed (+Tetra), Fast (-Tetra) and Fast (+Tetra) samples. All genes displayed are differentially expressed (p ≤ 0.05, log2FC ≥ 2 or ≤ –2). n = 2 mice per group. h. Bar graph showing the number of shared accessible regions between scATAC-seq and H3K27ac CUT&Tag datasets. “Common” refers to overlapping enhancer-like peaks. i. HOMER motif enrichment analysis showing -log10 p-values for transcription factor motifs associated with intestinal progenitor identity and injury response (IR) protection, alongside their corresponding gene expression levels from bulk RNA-seq in fed and fasted conditions and at 0-, 24-, 48-, and 72-hours post irradiation (hpi). j. Graphical representation of data shown in panel i for fed and 24 fasted mice. Y axis represents binned -log10 p-values of transcription factor motif activity. k. Graphical representation of data show in panel i for fed and fasted mice. mice at 0-, 24-, 48-, and 72-hours post irradiation (hpi). Y axis represents binned -log10 p-values of transcription factor motif activity.

Article Snippet: Tissues were blocked with Protein Block (Dako # X0909) and/or normal serum (Vector ImmPRESS # MP-7401), followed by overnight incubation with 1:200 dilution of Olfm4 antibody (CST # 39141S) at 4°C.

Techniques: Irradiation, Single Cell, Sequencing, Marker, RNA Sequencing, Gene Expression, Activity Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Wnt5a suppresses colorectal cancer progression via TGF-β/NOTUM/OLFM4 axis in patient-derived organoids

doi: 10.1186/s12964-025-02364-z

Figure Lengend Snippet: Wnt5a represses OLFM4 and ATOH1 expression in the microwells array-based organoid. ( A-H ) Organoids that express doxycycline (Dox)-inducible Wnt5a were seeded in microwell array and cultured for 7 days. ( A ) Bright-field images showing the development of an organoid in microwells over a period of 7 days. Scale bars: 100 μm. ( B ) Immunofluorescence images of organoids grown in microwell, these cells were labeled with Ki67 (green), EpCAM (red) and DAPI (blue). Scale bars: 100 μm. ( C ) Representative images of organoids treated with DMSO (left) or Dox (right). The black scale bars represent 100 μm; the white scale bars represent 300 μm. ( D ) Quantification of organoid sizes in ( C ). (E ) Immunofluorescence images of organoids grown in microwell, these cells were labeled with phospho-Smad2 (green), Wnt5a-Flag (red) and DAPI (blue). Scale bars: 100 μm. ( F ) RT–qPCR analysis of the expression of LGR5 and OLFM4 . The data are presented as mean ± SEM ( n = 3). ( G ) Immunofluorescence staining of organoids in the presence of DMS or Dox for OLFM4 (green), Wnt5a-Flag (red) and DAPI (blue). Scale bars: 100 μm. ( H - I ) RT-qPCR analysis of secretory markers (H: MUC2 , DEFA6 ) and ATOH1 ( I ). The data are presented as mean ± SEM ( n = 3)

Article Snippet: Antibodies to p-Smad2 (18338 S) and NOTUM (92654 S) were purchased from Cell Signaling Technology; antibody to Flag (PM020) was purchased from MBL; Antibody to Wnt5a (MA5-15502) was purchased from Thermo Fisher Scientific; Antibody to OLFM4 (MAB9218-100) was purchased from R&D Systems; Antibody to MUC2 (MA5-12345) was purchased from Invitrogen; Goat Anti-Rabbit IgG H&L (HRP, ab205718) was purchased from Abcam; Goat Anti-Mouse IgG, Light-Chain Specific Antibody (HRP Conjugate, 91196 S) was purchased from Cell Signaling Technology.

Techniques: Expressing, Cell Culture, Immunofluorescence, Labeling, Quantitative RT-PCR, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: Wnt5a suppresses colorectal cancer progression via TGF-β/NOTUM/OLFM4 axis in patient-derived organoids

doi: 10.1186/s12964-025-02364-z

Figure Lengend Snippet: Wnt5a-mediated inhibition of organoids growth requires NOTUM. ( A ) Organoids were treated with DMSO or Dox for 7days. RT–qPCR analysis of the expression of NOTUM and ATOH8 . The data are presented as mean ± SEM ( n = 3). ( B ) Protein association network for NOTUM using the STRING database. ( C ) Organoids were treated with DMSO or Dox for 7days. Western blot of NOTUM using anti-NOTUM. (D-E) Organoids were RT–qPCR ( D ) and Western blot ( E ) analysis of NOTUM expression in organoids treated with DMSO or Dox in the presence or absence of SB431542 for 7 days. The data are presented as mean ± SEM ( n = 3). ( F - G ) Organoids were treated with or without TGF-β (10 ng/mL) in the presence or absence of SB431542 for 7 days. Expression of NOTUM mRNA ( F ) and NOTUM levels were blotted with anti-NOTUM ( G ). The data are presented as mean ± SEM ( n = 3). ( H ) Representative images of organoids treated with DMSO or DOX in the presence or absence of ABC99 (500 nM) for 7 days. Scale bars: 100 μm. Quantification of organoid sizes in ( H ). ( I ) Organoids were treated with or without Dox in the presence or absence of ABC99 for 7 days. OLFM4 levels were blotted with anti-OLFM4

Article Snippet: Antibodies to p-Smad2 (18338 S) and NOTUM (92654 S) were purchased from Cell Signaling Technology; antibody to Flag (PM020) was purchased from MBL; Antibody to Wnt5a (MA5-15502) was purchased from Thermo Fisher Scientific; Antibody to OLFM4 (MAB9218-100) was purchased from R&D Systems; Antibody to MUC2 (MA5-12345) was purchased from Invitrogen; Goat Anti-Rabbit IgG H&L (HRP, ab205718) was purchased from Abcam; Goat Anti-Mouse IgG, Light-Chain Specific Antibody (HRP Conjugate, 91196 S) was purchased from Cell Signaling Technology.

Techniques: Inhibition, Quantitative RT-PCR, Expressing, Western Blot

Journal: Gut Microbes

Article Title: Time-restricted feeding promotes glucagon-like peptide-1 secretion and regulates appetite via tryptophan metabolism of gut Lactobacillus in pigs

doi: 10.1080/19490976.2025.2467185

Figure Lengend Snippet: Effect of feeding patterns on intestinal stem cell proliferation and differentiation. (a) The mRNA expressions of epithelial cell marker genes in the ileum ( n = 7). (b) The mRNA expressions of epithelial cell marker genes in the colon ( n = 7). (c) Immunofluorescence staining of Olfm4, Muc2 and ChgA in pig colonic tissue ( n = 6). (d) The protein expression of ChgA ( n = 3) in the colon. (e) The mRNA expressions of enteroendocrine cell marker genes ( n = 7) in the colon. (f) The mRNA expressions of enteroendocrine cell transcription factors ( n = 7) in the colon. (g) The protein expression of Notch1 ( n = 3) in the colon. Data are presented as the mean ± SEM. Different letters denote statistically significant differences among the groups ( p < 0.05).

Article Snippet: Tissue sections were then subjected to blocking and incubated with primary antibodies targeting Olfm4 (1:100 diluted; Cell Signaling Technology, #14369), Muc2 (1:100 diluted; ABclonal, A14659), and ChgA (1:100 diluted; ABclonal, A9576).

Techniques: Marker, Immunofluorescence, Staining, Expressing