recombinant mouse tgf β  (R&D Systems)

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: Primers used to detect mouse genes

Article Snippet: To measure accelerated HSC activation in vitro, mHSCs were cultured for 7 days and challenged with 10 ng/mL of recombinant mouse TGF –β (R&D Systems #7666-MB) as a positive control of HSC activation, or recombinant mouse Wnt3a (R&D Systems #1324-WN) or Wnt5a (R&D Systems #645-WN), DKK-1 (R&D Systems #5897-DK) on day 5.

Techniques:

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: Exogenous Wnt3a and Wnt5a accelerate the spontaneous activation of mouse hepatic stellate cells. A ) Primary mouse hepatic stellate cells were isolated then cultured in the presence or absence of 10 ng/mL of exogenous TGF-β, Wnt3a, and Wnt5a from day 5 to day 7. B ) Expression of col1a1 mRNA was detected in mHSCs by qRT-PCR. C ) mHSC lysates were prepared and proteins separated by SDS-PAGE. α-SMA and HSC70 (loading control) were measured by Western blot. D ) β-catenin and Desmin expression was analyzed by immunofluorescence in cells following TGF-β, Wnt3a, and Wnt5a treatment. Red arrows highlight nuclear localization of β-catenin. E ) β-catenin expression and nuclear localization, and Desmin expression by immunofluorescence was quantified using ImageJ software. F ) Expression of axin2, ctgf, cyclind1, mmp9, mmp13, and timp1 mRNA and G ) wnt3a and wnt5a mRNA was detected by qRT-PCR. N = 5–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: To measure accelerated HSC activation in vitro, mHSCs were cultured for 7 days and challenged with 10 ng/mL of recombinant mouse TGF –β (R&D Systems #7666-MB) as a positive control of HSC activation, or recombinant mouse Wnt3a (R&D Systems #1324-WN) or Wnt5a (R&D Systems #645-WN), DKK-1 (R&D Systems #5897-DK) on day 5.

Techniques: Activation Assay, Isolation, Cell Culture, Expressing, Quantitative RT-PCR, SDS Page, Control, Western Blot, Immunofluorescence, Software

Journal: Cell Biology and Toxicology

Article Title: Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury

doi: 10.1007/s10565-024-09956-4

Figure Lengend Snippet: HSC activation, via LRP6, occurs in a canonical Wnt-dependent manner. A ) Primary mouse HSCs were then cultured in the presence or absence of TGF-β, Wnt3a, and Wnt5a and the LRP-6 antagonist DKK-1 (10 ng/mL) from day 5 to day 7. B ) Expression of col1a1 mRNA was determined by qRT-PCR. C ) mHSC lysates were prepared and proteins separated by SDS-PAGE. α-SMA expression and HSC70 (loading control) were measured by Western Blot. α-SMA was normalized to HSC70 as a loading control and quantified using ImageJ software. D ) Expression of axin2, mmp9, and mmp13 mRNA; and E ) wnt3a and wnt5a mRNA was determined in mHSCs by qRT-PCR. Data are from three independent experiments. N = 3–6 per group. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Article Snippet: To measure accelerated HSC activation in vitro, mHSCs were cultured for 7 days and challenged with 10 ng/mL of recombinant mouse TGF –β (R&D Systems #7666-MB) as a positive control of HSC activation, or recombinant mouse Wnt3a (R&D Systems #1324-WN) or Wnt5a (R&D Systems #645-WN), DKK-1 (R&D Systems #5897-DK) on day 5.

Techniques: Activation Assay, Cell Culture, Expressing, Quantitative RT-PCR, SDS Page, Control, Western Blot, Software