Fig 1: MMP-2 activity is reduced during the sustained depressive-like state. (a) In-gel zymography assays (Zy) were performed at 72 h and 8 weeks after social defeat. (b) No difference in MMP-2 or MMP-9 activity was present shortly after social defeat; n = 6 animals per group. (c) At week 8 post-defeat, decreased MMP-2 activity was found in the SDPS group, without MMP-9 regulation; n = 6 animals (control); n = 4 (SDPS). *P < 0.05. Data are expressed as mean ± SEM. (d) Examples with equal loading showing the mature form of MMP-2 and MMP-9 (white bands), and the position of the 75 kDa marker band for each time point. Full gels are available in Supplementary Fig. 6 online.
Fig 2: Increase of serum and CSF MMP9 and albumin in idiopathic epileptic dogs. (A) Serum gel zymography results of control and IE dogs show different expression patterns of subtypes of MMP9 proteins (black arrows indicate active form) in some of the samples from IE dogs. (B) Graph showing quantification of serum MMP9 complex zymography results of control (n = 5) and IE (n = 21) dogs, with significantly higher levels in the IE group (P = .04, marked by an asterisk). (C) CSF gel zymography results of control and IE dogs showing different expression patterns of subtypes of MMP9 proteins (black arrows indicate active form) in some of the samples of IE dogs. (D) Graph showing quantification of CSF MMP9 complex zymography results of control (n = 4) and IE (n = 16) dogs, with a significantly higher levels in the IE group (P = .02, marked by an asterisk). (E) Albumin quota ratio was significantly higher (P = .04) in the IE group (n = 18) than in control dogs (n = 6). (F) Levels of BBB score, MMP9 and AQ of 1 IE dog at 2 time points (2 months apart) showing increases in all variables. (G) Serum MMP9 gel zymography results of the IE dog shown in panel F at the same 2 time points.
Fig 3: MMP9 levels in the piriform cortex of dogs. (A) Bar plots showing MMP9 levels as percentage of pixels with fluorescence signal in each image (left) and as fluorescence intensity (right) in the piriform cortex of control (n = 4) and IE dogs (n = 5). Higher levels of MMP9 were measured in IE dogs (P = .04). (B) Microscopy images of sections from the piriform cortex of representative control (a) and IE dogs (b) stained for MMP9 (green). Increased MMP9 expression is seen in the epileptic dog (scale bar, 20 μm).
Fig 4: Co‐localization of albumin and MMP9 in neurons in the piriform cortex of IE dogs. Microscopic images of coronal sections from the piriform cortex of control (upper image) and IE dogs stained for astrocytic marker GFAP (red), MMP9 (green), albumin (white), and DAPI nuclear staining (blue). In the epileptic dog, albumin uptake and increased MMP9 expression co‐localized to nonastrocyte cells with large nuclei. Scale bars, 20 μm.
Fig 5: Sex-mismatched secretomic environments differentially influence macrophage behavior in 3D culture.Exposure to a sex-mismatched secretome is associated with a trend toward increased proliferation in RAW 264.7 macrophages, while showing a negative trend in proliferation for J774A.1 cells. Additionally, sex-mismatched conditions elevate MMP9 expression in RAW 264.7 macrophages. (A) Ki67+ over DAPI+ ratios based on immunocytochemistry imaging for RAW 264.7 (striped) and J774A.1 (dotted) in sex-matched and sex mis-matched environments. (B) MMP9 expression via zymography for RAW 264.7 (striped) and J774A.1 (dotted) in sex-matched and sex mis-matched environments. For figure – BM refers to basal media, RM (green) refers to RAW 264.7 conditioned media, JM (pink) refers to J774A.1 conditioned media. One-way ANOVA and post hoc Tukey statistical analysis conducted (*p<.05,**p<.01,***p<.001).
Supplier Page from Abcam for Recombinant Mouse MMP9 protein