rabbit anti-mlck  (Servicebio Inc)

Journal: iScience

Article Title: Trifluoperazine regulates blood-brain barrier permeability via the MLCK/p-MLC pathway to promote ischemic stroke recovery

doi: 10.1016/j.isci.2024.109156

Figure Lengend Snippet: TFP regulates BBB permeability via the MLCK/p-MLC signaling pathway and contributes to neurological recovery (A) Western blotting analysis of MLCK, p-MLC, and MLC in mice after I/R in the MCAO, MCAO+TFP, MCAO+TFP+LV CON, and MCAO+TFP+LV MLCK-OE groups. (B) Quantitative analysis of the expression levels of MLCK and p-MLC in (A) (n = 3 per group). (C) Representative immunofluorescence images of p-MLC (red), CD31 (green), and DAPI (blue) staining of mouse brain tissues after I/R in the sham, MCAO, MCAO+TFP, MCAO+TFP+LV CON, and MCAO+TFP+LV MLCK-OE groups (scale bars, 50 μm). (D) Quantitative analysis of CD31 + p-MLC + cells as a percentage of CD31 + cells in (D) (n = 6 per group). Data are represented as mean ± SEM. ns, p > 0.05, ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: The data obtained were validated using rabbit anti-CaM antibody (1:1000, Abcam, Cambridge, MA, USA), rabbit anti-Claudin-5 antibody (1:1000, Abcam, Cambridge, MA, USA), rabbit anti-Occludin antibody (1:1000, CST, Danvers, MA, USA), rabbit anti-ZO-1 antibody (1:1000, Invitrogen, Carlsbad, CA, USA), rabbit anti-MLCK antibody (1:1000, Proteintech, Wuhan, China), rabbit anti-MLC antibody (1:1000, Abcam, Cambridge, MA, USA), and rabbit anti-p-MLC antibody (1:1000, CST, Danvers, MA, USA).

Techniques: Permeability, Western Blot, Expressing, Immunofluorescence, Staining

Journal: iScience

Article Title: Trifluoperazine regulates blood-brain barrier permeability via the MLCK/p-MLC pathway to promote ischemic stroke recovery

doi: 10.1016/j.isci.2024.109156

Figure Lengend Snippet:

Article Snippet: The data obtained were validated using rabbit anti-CaM antibody (1:1000, Abcam, Cambridge, MA, USA), rabbit anti-Claudin-5 antibody (1:1000, Abcam, Cambridge, MA, USA), rabbit anti-Occludin antibody (1:1000, CST, Danvers, MA, USA), rabbit anti-ZO-1 antibody (1:1000, Invitrogen, Carlsbad, CA, USA), rabbit anti-MLCK antibody (1:1000, Proteintech, Wuhan, China), rabbit anti-MLC antibody (1:1000, Abcam, Cambridge, MA, USA), and rabbit anti-p-MLC antibody (1:1000, CST, Danvers, MA, USA).

Techniques: Recombinant, Lysis, Software

Journal: iScience

Article Title: Trifluoperazine regulates blood-brain barrier permeability via the MLCK/p-MLC pathway to promote ischemic stroke recovery

doi: 10.1016/j.isci.2024.109156

Figure Lengend Snippet: TFP regulates BBB permeability via the MLCK/p-MLC signaling pathway and contributes to neurological recovery (A) Western blotting analysis of MLCK, p-MLC, and MLC in mice after I/R in the MCAO, MCAO+TFP, MCAO+TFP+LV CON, and MCAO+TFP+LV MLCK-OE groups. (B) Quantitative analysis of the expression levels of MLCK and p-MLC in (A) (n = 3 per group). (C) Representative immunofluorescence images of p-MLC (red), CD31 (green), and DAPI (blue) staining of mouse brain tissues after I/R in the sham, MCAO, MCAO+TFP, MCAO+TFP+LV CON, and MCAO+TFP+LV MLCK-OE groups (scale bars, 50 μm). (D) Quantitative analysis of CD31 + p-MLC + cells as a percentage of CD31 + cells in (D) (n = 6 per group). Data are represented as mean ± SEM. ns, p > 0.05, ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: We used the following primary antibodies and secondary antibodies at the indicated dilutions: goat anti-CD31 Alexa Fluor 488-conjugated antibody (1:400, R&D, Minneapolis, MN, USA), rabbit anti-Claudin-5 (1:100, Abcam, Cambridge, MA, USA), rabbit anti-Occludin (1:100, CST, Danvers, MA, USA), rabbit anti- zonula occludens (ZO)-1 (1:200, Invitrogen, Carlsbad, CA, USA), rabbit anti-MLCK (1:200, Abcam, Cambridge, MA, USA), rabbit anti-p-MLC (1:50, CST, Danvers, MA, USA), and anti-rabbit Alexa Fluor 488 or Alexa Fluor 594 (1:400, Abcam, Cambridge, MA, USA).

Techniques: Permeability, Western Blot, Expressing, Immunofluorescence, Staining

Journal: iScience

Article Title: Trifluoperazine regulates blood-brain barrier permeability via the MLCK/p-MLC pathway to promote ischemic stroke recovery

doi: 10.1016/j.isci.2024.109156

Figure Lengend Snippet:

Article Snippet: We used the following primary antibodies and secondary antibodies at the indicated dilutions: goat anti-CD31 Alexa Fluor 488-conjugated antibody (1:400, R&D, Minneapolis, MN, USA), rabbit anti-Claudin-5 (1:100, Abcam, Cambridge, MA, USA), rabbit anti-Occludin (1:100, CST, Danvers, MA, USA), rabbit anti- zonula occludens (ZO)-1 (1:200, Invitrogen, Carlsbad, CA, USA), rabbit anti-MLCK (1:200, Abcam, Cambridge, MA, USA), rabbit anti-p-MLC (1:50, CST, Danvers, MA, USA), and anti-rabbit Alexa Fluor 488 or Alexa Fluor 594 (1:400, Abcam, Cambridge, MA, USA).

Techniques: Recombinant, Lysis, Software

Journal: Journal of Neuroinflammation

Article Title: Inhibition of UTX/KDM6A improves recovery of spinal cord injury by attenuating BSCB permeability and macrophage infiltration through the MLCK/p-MLC pathway

doi: 10.1186/s12974-023-02936-1

Figure Lengend Snippet: Conditional UTX knockout in ECs regulates BSCB permeability following SCI via the MLCK/p-MLC pathway, promoting neurological recovery. a Western blotting analysis of MLCK, p-MLC, MLC in UTX f/f , UTX −/− , UTX −/− + LV CON, and UTX −/− + LV MLCK-OE mice at SCI 3d. b , c Quantitative analysis of the expression levels of MLCK and p-MLC in a . n = 3 per group. d Representative immunofluorescence images of p-MLC (red) and CD31 (green), DAPI (blue) staining of injured epicenter of spinal cord in UTX f/f , UTX −/− , UTX −/− + LV CON, and UTX −/− + LV MLCK-OE mice at SCI 3d. Scale bars, 50 μm. e Quantitative analysis of CD31 + p-MLC + cells as a percentage of CD31 + cells in d , n = 6 per group. f , g Distribution of the BMS scores and BMS sub-scores in UTX f/f , UTX −/− , UTX −/− + LV CON, and UTX −/− + LV MLCK-OE mice at pre-surgery and after SCI throughout the 56-day period. n = 12 per group. h Representative images of motor-evoked potential (MEP) in UTX f/f , UTX −/− , UTX −/− + LV CON, and UTX −/− + LV MLCK-OE mice at 56-day post-SCI. i Quantitative evaluation of the amplitude of MEPs in h . n = 6 per group. Data are represented as mean ± SEM. ns P > 0.05, * P < 0.05, ** P < 0.01

Article Snippet: We utilized the following primary antibodies, secondary antibodies and dilutions: goat anti-CD31 Alexa Fluor 488-conjugated antibody (1:200, R&D, USA), rabbit anti-UTX (1:100, MilliporeSigma, USA), rabbit anti-Claudin5 (1:100, Abcam, USA), rabbit anti-Occludin (1:100, CST, USA), rabbit anti-Zona occludens 1(ZO-1) (1:200, Invitrogen, USA), rat anti-F4/80(1:200, Abcam, USA), rabbit MLCK (1:200, Abcam, USA), rabbit anti-p-MLC (1:50, CST, USA), and anti-goat Alexa Fluor 488 or anti-rabbit / anti-rat Alexa Fluor 594 (1: 400, Abcam, USA).

Techniques: Knock-Out, Permeability, Western Blot, Expressing, Immunofluorescence, Staining