Anti-LGR5 antibody [EPR3065Y]

• WB

Unknown

Western blot - Anti-LGR5 antibody [EPR3065Y] (AB75850)

Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-LGR5 antibody [EPR3065Y] (ab75850) at 1/1000 dilution

All lanes:

Human skeletal muscle lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 99 kDa

Observed band size: 100 kDa

false

• WB

Supplier Data

Western blot - Anti-LGR5 antibody [EPR3065Y] (AB75850)

We are unsure about the identity of the additional band.

Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-LGR5 antibody [EPR3065Y] (ab75850) at 1/1000 dilution

Lane 1:

Mouse skeletal muscle lysate at 20 µg

Lane 2:

Rat skeletal muscle lysate at 20 µg

Lane 3:

Mouse brain lysate at 20 µg

Lane 4:

Rat brain lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 99 kDa

Observed band size: 100 kDa

false

• WB

Unknown

Western blot - Anti-LGR5 antibody [EPR3065Y] (AB75850)

All lanes:

Western blot - Anti-LGR5 antibody [EPR3065Y] (ab75850) at 1/1000 dilution

All lanes:

Fetal skeletal muscle tissue lysate at 10 µg

Secondary

All lanes:

HRP-labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 99 kDa

Observed band size: 100 kDa

false

• WB

CiteAb

Western blot - Anti-LGR5 antibody [EPR3065Y] (AB75850)

Western Blotting using Anti-LGR5 antibody [EPR3065Y], ab75850. Publication image from Shui, J. W. et al., 2022, Nat Commun, 35169117. Legend direct from paper.

IL-18-Akt-Tcf4 signalling upregulates Lgr5+ stem cells.a–c Flow cytometry analysis of CD24-/low ileum crypts, isolated from the indicated uninfected or AIEC-infected mice at d6, for Lgr5+ stem cells. An isotype antibody was included to indicate the specificity of the Lgr5 antibody. d Immunofluorescence analysis of ileum crypts for Olfm4+ stem cells in the indicated uninfected or AIEC-infected mice at d6. Crypt base columnar (CBC, outlined with a solid line) stem cell and the above transit-amplifying (TA, outlined with a dashed line) compartments are indicated. e Flow cytometry analysis of IL-18-stimulated ileum organoids, derived from the indicated mice, for Lgr5+ stem cells. f Chromatin immunoprecipitation (ChIP) and PCR analyses of IL-22 or IL-18-stimulated wild-type ileum organoids for the binding of Tcf4 to the mouse Lgr5 promoter regions (P1~P6). Fold induction is calculated based on PCR signal strength of Tcf4-bound Lgr5 promoter P1 region before and after IL-22 or IL-18 stimulation. g Western blot and flow cytometry analyses of IL-18/Akt inhibitor-stimulated CMT93 cells for Akt phosphorylation and Lgr5. Quantification and the ratio (pAkt to total Akt, Lgr5 to β-actin) of protein bands are indicated. h ChIP and PCR analyses of IL-18/Akt inhibitor-stimulated CMT93 cells for the binding of Tcf4 to the mouse Lgr5 promoter regions (P1 and P5). Fold induction is calculated based on PCR signal strength of Tcf4-bound Lgr5 promoter P1 region before and after stimulation. Each symbol in bar graphs represents a well of CMT93 cell culture (g), an ileum crypt sample (a–c) or organoid culture (e, f) derived from one mouse. Data shown are representative (d, f–h) or combined (a–c, e) results from two independent reproducible experiments. Statistical significance is indicated using unpaired two-tailed t test (a–c), One-way ANOVA with Sidak’s multiple comparisons test (f, g), or Two-way ANOVA with Tukey’s multiple comparisons test (e). Data are presented as mean ± SD. Source data are provided as a Source Data file.

false

• WB

CiteAb

Western blot - Anti-LGR5 antibody [EPR3065Y] (AB75850)

Western Blotting using Anti-LGR5 antibody [EPR3065Y], ab75850. Publication image from Lin, Y. et al., 2017, Nat Commun, 29079780. Legend direct from paper.

Knocking down of Lgr5 expression decreases Lgr5+ cells’ stem cell properties and increases liver fibrosis. a Lgr5 was knocked down by Ad-Lgr5 shRNA. Lgr5+ liver stem cells were isolated and infected with Ad-Lgr5 shRNA, or nonsense control shRNA Ad-Ctrl shRNA, western blot, and qRT-PCR were used to analyze the proteins level (left) and the mRNA level (right) of Lgr5. Actin was used as a loading control. b Lgr5+ liver stem cells failed to form an organoid when Lgr5 was downregulated by Ad-Lgr5 shRNA. Lgr5+ liver stem cells were isolated and infected with Ad-Lgr5 shRNA or Ad-Ctrl shRNA, and cultured in stem cells medium in Matrigel, and the number and sizes of the liver organoid were measured at day 14. b–e Ad-Lgr5 shRNA increased CCL4-induced liver fibrosis. As shown in Supplementary Fig. 10a, Lgr5-GFP mice infected with Ad-Lgr5 shRNA or Ad-Ctrl shRNA were administered with CCL4 (2 ml/kg i.p.) twice/week for 6 weeks, respectively. The representative histology of H&E and Sirius Red, and the quantification of positive-staining areas is measured by Image J software (c). Serum levels of ALT and AST were measured (d, e). For a, d, e, triplicates for each condition were analyzed. The results are shown as mean ± s.d. of three independent experiments. *p < 0.05. For b, the results are shown as mean ± s.d. of three independent experiments. **p < 0.01. For c, scale bars, 200 µm, the results are shown as mean ± s.d. of five independent sections taken randomly per mice and a total of 50 tissue specimens in each group (n = 10 mice), *p < 0.05

false