primary human brain microvascular endothelial cells (hbmecs; acbri376)  (Cell Systems Corporation)

Journal: Molecular neurobiology

Article Title: Recombinant FGF21 protects against blood-brain barrier leakage through Nrf2 upregulation in type 2 diabetes mice

doi: 10.1007/s12035-018-1234-2

Figure Lengend Snippet: Cultured HBMEC monolayer treated with HG-IL1β was used to mimic BBB injury under diabetic conditions. The monolayer was then treated with rFGF21 and the permeability was measured. (A) IL-1β protein level in diabetic mice brain measured by Western blot (* p<0.05 vs. db/+, n=4); (B) Viability of cultured HBMEC after treatment with HG-IL1β and rFGF21; (C) Relative HBMEC monolayer permeability after treatment with HG-IL1β and rFGF21 (* p<0.05 vs. norm; # p<0.05 vs. HG-IL1β; n=6); (D) Representative western blot images of junction protein expression in cultured HBMEC after HG-IL1β and rFGF21 treatment; (E, F, G) Quantification of junction protein ZO-1, VE-cadherin and Occludin in cultured HBMEC (*p<0.05 vs. norm; # p<0.05 vs. HG-IL1β; n=6); (H) Immunocytochemistry of ZO-1 and VE-cadherin in cultured HBMEC after HG-IL1β and rFGF21 treatment.

Article Snippet: Primary HBMEC was obtained from Cell Systems Corporation (ACBRI376, Kirkland, WA), and cultured in complete growth media EBM-2 containing supplements growth factors (Lonza, Walkersville, MD).

Techniques: Cell Culture, Permeability, Western Blot, Expressing, Immunocytochemistry

Journal: Molecular neurobiology

Article Title: Recombinant FGF21 protects against blood-brain barrier leakage through Nrf2 upregulation in type 2 diabetes mice

doi: 10.1007/s12035-018-1234-2

Figure Lengend Snippet: The FGFR1 inhibitor, PD173074, was used to examine the roles of FGFR1 in BBB protection effect by FGF21 using cultured HBMEC monolayer. (A) Relative permeability of HBMEC monolayer after HG-IL1β and FGF21 treatment with or without PD173074; (B) Representative Western Blot images to measure FGFR1, pFGFR1 and β-Klotho in cultured HBMEC after HG-IL1β and FGF21 treatment with or without PD173074; (C) Quantification of pFGFR1 in cultured HBMEC. (* p<0.05 vs. HG-IL1β; #p<0.05 vs. HG-IL1β+F21; n=4).

Article Snippet: Primary HBMEC was obtained from Cell Systems Corporation (ACBRI376, Kirkland, WA), and cultured in complete growth media EBM-2 containing supplements growth factors (Lonza, Walkersville, MD).

Techniques: Cell Culture, Permeability, Western Blot

Journal: Molecular neurobiology

Article Title: Recombinant FGF21 protects against blood-brain barrier leakage through Nrf2 upregulation in type 2 diabetes mice

doi: 10.1007/s12035-018-1234-2

Figure Lengend Snippet: The involvement of Nrf2 in FGF21 protection for BBB was examined using db/db mice and cultured HBMEC monolayer. (A) Nrf2 and pNrf2 protein levels measured by Western blot in nuclear extract of db/db mice brain after rFGF21 treatment (* p<0.05 vs. db/+; # p<0.05 vs. db/db; n=4); (B) Nrf2 and pNrf2 protein levels measured by Western blot in cultured HBMEC after HG-IL1β and rFGF21 treatment (* p<0.05 vs. norm; # p<0.05 vs. HG-IL1β; n=3). (C) Relative HBMEC monolayer permeability after treatment with HG-IL1β and rFGF21, with or without Trigonelline, the Nrf2 inhibitor (* p<0.05 vs. norm; # p<0.05 vs. HG-IL1β; & p<0.05 vs. HG-IL1β+F21; n=4). The roles of Nrf2-Keap1 interaction in BBB protection by FGF21 was further investigated using db/db mice and cultured HBMEC. (D) Representative Western blot images and quantification for Keap1 protein in db/db mice brain after FGF21 treatment; (E) Representative Western blot images and quantification for Nrf2 and pNrf2 detection after Co-IP with Keap1 antibody using the brain lysate of db/db mice after rFGF21 treatment (* p<0.05 vs. db/+; # p<0.05 vs. db/db; n=4). Keap1 and Nrf2 interaction was also measured in cultured HBMEC after HG-ΙL1β and rFGF21 treatment. (F) Keap1 Western blot and quantification in cultured HBMCE; (G) Co-IP was performed using Keap1 antibody with HBMCE lysate, and pNrf2 in the precipitate was assessed using Western blot (* p<0.05 vs. norm; # p<0.05 vs. HG- IL1β; n=3). (H) Representative Western blot images of Smad2, p-Smad2 and Snail in db/db mice after rFGF21 treatment.

Article Snippet: Primary HBMEC was obtained from Cell Systems Corporation (ACBRI376, Kirkland, WA), and cultured in complete growth media EBM-2 containing supplements growth factors (Lonza, Walkersville, MD).

Techniques: Cell Culture, Western Blot, Permeability, Co-Immunoprecipitation Assay