bk037  (Cytoskeleton Inc)

Journal: Molecular Medicine Reports

Article Title: Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis

doi: 10.3892/mmr.2026.13813

Figure Lengend Snippet: FRA1 expression in MRL/lpr mouse kidneys. Representative H&E staining of kidney sections from 20-week-old (A) MRL/lpr mice and (B) MRL/MPJ controls, showing tubulointerstitial inflammation and damage in MRL/lpr mice. MRL/lpr mice exhibit diffuse and extensive tubulointerstitial inflammation and structural damage. (C-H) FRA1 expression in mouse kidney sections. (C, E and G) Three independent mice from the MRL/lpr group, all demonstrating elevated FRA1 levels in the tubular epithelium. (D, F and H) Three independent mice from the control group, displaying baseline FRA1 immunoreactivity. (I) Quantification of FRA1-positive area from IHC images. (J) Western blot analysis of FRA1 protein levels from MRL/lpr and control mice. (K) Densitometric quantification of FRA1 bands normalized to β-actin. Scale bar, 50 µm; Data are plotted as the mean ± SEM; n=3 per group; group comparisons were performed using two-sided Welch's t-tests; ***P<0.001.

Article Snippet: The primary antibodies used in the present study included: β-actin (1:5,000; cat. no. bs-0061R; BIOSS), FRA1 (1:1,000; cat. no. A5372; ABclonal Biotech Co., Ltd.), IL-1β (1:1,000; cat. no. HA601002 ; clone no. A7F9; HUABIO), IL-8 (1:1,000; cat. no. ab289967; clone no. EPR26511-74; Abcam), RANTES (1:1,000; cat. no. 36467; CST Biological Reagents Co., Ltd.), IL-6 (1:2,000; cat. no. DF6087; Affinity Biosciences, Ltd.), MCP-1 (1:2,000; cat. no. A7277; ABclonal Biotech Co., Ltd.), TNF-α (1:2,000; cat. no. 17590-1-AP; Proteintech Group, Inc.), and TGF-β (1:2,000; cat. no. 81746-2-RR; clone no. 230544B7; Proteintech Group, Inc.).

Techniques: Expressing, Staining, Control, Western Blot

Journal: Molecular Medicine Reports

Article Title: Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis

doi: 10.3892/mmr.2026.13813

Figure Lengend Snippet: Effect of FRA1 on inflammatory cytokine expression in HK-2 cells. (A) Representative western blots of FRA1 and inflammatory cytokines (IL-1β, IL-6, and IL-8) in HK-2 cells 144 h after transduction with FRA1-OE, FRA1-shRNA or their corresponding controls. (B) Representative western blots of MCP-1, RANTES, TGF-β and TNF-α in HK-2 cells following the same transduction conditions. Densitometric semi-quantification of protein bands normalized to β-actin for: (C) FRA1, (D) IL-1β, (E) IL-6, (F) IL-8, (G) MCP-1, (H) RANTES, (I) TGF-β and (J) TNF-α. Data are presented as the mean ± SEM; n=3 per group; comparisons among subgroups were assessed using the two-sided Kruskal-Wallis test; *P<0.05, **P<0.01 and ***P<0.001. OE, over expression; ns, not significant; sh, short hairpin.

Article Snippet: The primary antibodies used in the present study included: β-actin (1:5,000; cat. no. bs-0061R; BIOSS), FRA1 (1:1,000; cat. no. A5372; ABclonal Biotech Co., Ltd.), IL-1β (1:1,000; cat. no. HA601002 ; clone no. A7F9; HUABIO), IL-8 (1:1,000; cat. no. ab289967; clone no. EPR26511-74; Abcam), RANTES (1:1,000; cat. no. 36467; CST Biological Reagents Co., Ltd.), IL-6 (1:2,000; cat. no. DF6087; Affinity Biosciences, Ltd.), MCP-1 (1:2,000; cat. no. A7277; ABclonal Biotech Co., Ltd.), TNF-α (1:2,000; cat. no. 17590-1-AP; Proteintech Group, Inc.), and TGF-β (1:2,000; cat. no. 81746-2-RR; clone no. 230544B7; Proteintech Group, Inc.).

Techniques: Expressing, Western Blot, Transduction, shRNA, Over Expression

Journal: Redox Biology

Article Title: Myo1f regulates monocyte adhesion and contributes to atherosclerosis via MRTFA-dependent ITGB2 expression

doi: 10.1016/j.redox.2026.104049

Figure Lengend Snippet: Cytoskeletal polymerization promotes ITGB2 transcription and MRTFA nuclear translocation. A, Western blot analysis of ITGB2 expression after increasing gradient of G-actin polymerization inhibitor LAT-A under oxLDL stimulation (n = 3). B, The adhesion of THP-1 cells to HUVECs after treatment with LAT-A (2 μM) was observed under a fluorescence microscope (scale bar, 50 μm). Adherent cell numbers were calculated from 3 biological replicate images, and data are shown as fold change (ratio to DMSO). C, After overexpressing Myo1f and treating THP-1 cells with or without LAT-A, Western blot analysis was performed on ITGB2 expression under oxLDL stimulation (n = 3). D, Nuclear translocation of MRTFA after LAT-A treatment based on oxLDL stimulation by dissociated cytoplasmic and nuclear analysis (n = 3). E, Representative colocalization immunofluorescence micrographs of phalloidin-labeled F-actin (red) and Myo1f (green) staining in THP-1 cells. The scan line graph represents the F-actin and Myo1f staining intensity along the white straight line (scale bar, 20 μm; n = 3). F, By isolating F-actin and G-actin in THP-1 cells exposed to oxLDL with or without depletion of Myo1f, and quantifying their ratios by immunoblotting (n = 3). G, Effects of knockdown of Myo1f on MRTFA and actin binding in THP-1 cells. MRTFA antibody was used to co-immunoprecipitate actin in THP-1 cells with or without Myo1f knockdown treatment and stimulation with oxLDL (n = 3). Data were presented as the mean ± SEM. Student's t-test (unpaired, two-tailed) for F and G . One-way ANOVA followed by Tukey's multiple comparisons test for A-D . Each P value is displayed in the image.

Article Snippet: The F-actin to G-actin ratio was determined following the manufacturer's instructions (BK037, Cytoskeleton).

Techniques: Translocation Assay, Western Blot, Expressing, Fluorescence, Microscopy, Immunofluorescence, Labeling, Staining, Knockdown, Binding Assay, Two Tailed Test