Journal: Molecular pain

Article Title: Activation and expression of μ-calpain in dorsal root contributes to RTX-induced mechanical allodynia.

doi: 10.1177/1744806917719169

Figure Lengend Snippet: Figure 1. Quantitative analysis of the expression and activation of m-calpain in L4–L6 dorsal root. (a) and (b) Representative gel images show the protein level of m-calpain, a II-spectrin, and a II-spectrin breakdown product (spectrin BD) in L4–L6 dorsal root obtained from vehicle group and RTX group rats. b-actin was used as a loading control. (c) Summary data show the protein level of m-calpain in L4–L6 dorsal root. (d) Summary data show the percentage of a II-spectrin breakdown product in the total of a II-spectrin in L4–L6 dorsal root. The calpain activity was estimated by evaluating the protein level of a II-spectrin breakdown product (150 kDa) using Western blotting. Data are expressed as means SEM (n ¼ 8 rats in each group). *P < 0.05, compared with the vehicle group. *P < 0.05, compared with the RTX group.

Article Snippet: The membrane was incubated with primary antibodies of rabbit antim-calpain antibody (1:1000, Cell Signaling Technology, Boston, USA); mouse anti-MBP antibody (1:20000, Novus Biological Littleton, CO, USA); mouse anti-a II-spectrin antibody (1:4000, Endo Life Sciences, Farmingdale, NY, USA); or mouse anti-b-actin antibody (1:5000; Santa Cruz, Dallas, TX, USA) at 4 C overnight.

Techniques: Expressing, Activation Assay, Control, Activity Assay, Western Blot

Journal: Molecular pain

Article Title: Activation and expression of μ-calpain in dorsal root contributes to RTX-induced mechanical allodynia.

doi: 10.1177/1744806917719169

Figure Lengend Snippet: Figure 4. The effect of calpain inhibitor MDL 28170 on RTX induced calpain activation and degradation of myelin basic protein (MBP) in L4–L6 dorsal root. (a) to (c) Representative gel images show the protein level of m-calpain, a II-spectrin, a II-spectrin breakdown product (spectrin BD), and 21.5- and 18.5-kDa MBP isoform in L4–L6 dorsal root obtained from vehicle, RTX 6 W, MDL 28170 (MDL), and PEG400/DMSO (P/D) rats. b-actin was used as a loading control. (d), (g), (f) Summary data show the protein level of m-calpain and 21.5- and 18.5-kDa MBP isoform in L4–L6 dorsal root. (e) Summary data show the percentage of a II-spectrin breakdown product in the total of a II-spectrin in L4–L6 dorsal root. Data are expressed as means SEM (n ¼ 6 rats in each group). *P < 0.05, compared with the vehicle group. #P < 0.05, compared with the PEG400/DMSO group.

Article Snippet: The membrane was incubated with primary antibodies of rabbit antim-calpain antibody (1:1000, Cell Signaling Technology, Boston, USA); mouse anti-MBP antibody (1:20000, Novus Biological Littleton, CO, USA); mouse anti-a II-spectrin antibody (1:4000, Endo Life Sciences, Farmingdale, NY, USA); or mouse anti-b-actin antibody (1:5000; Santa Cruz, Dallas, TX, USA) at 4 C overnight.

Techniques: Activation Assay, Control