Journal: The Journal of Cell Biology

Article Title: Spectrin coordinates cell shape and signaling essential for epidermal differentiation

doi: 10.1083/jcb.202502071

Figure Lengend Snippet: Spectrin determines junctional actomyosin network structure and stability. (a) Newborn epidermal whole-mount immunofluorescence analysis for phalloidin (F-actin) and non-muscle myosin heavy chain IIa (myosin-IIa). Minimal max. projections of the denoted layers are shown. (b) Dorsal skin sections from E17.5 wild-type embryos treated with DMSO or Y27632 (40 µM) immunolabeled for αII-spectrin. (c) Quantification of basal (left graph) and suprabasal (right graph) layer αII-spectrin intensity from data shown in b. Data are the mean ± SD of 30 ROI from n = 3 embryos per condition. Bars: mean normalized intensity; dots: microscopy fields. *P ≤ 0.05; NS: P = 0.3876 with Kolmogorov–Smirnov. (d) Immunofluorescence analysis for αII-spectrin, F-actin, and non-muscle myosin heavy chain IIa (myosin-IIa) (48 h high Ca 2+ ) in Ctr and αII-spectrin–deficient (KO) cells. (e) Immunofluorescence analysis for αII-spectrin and F-actin (48 h high Ca 2+ ) upon αII-spectrin ( Sptan1 ) knockdown and treatment with either DMSO or low-dose blebbistatin (5 µM). Representative images of n = 3 biological replicates each. (f) Immunofluorescence analysis for F-actin (48 h high Ca 2+ ) upon αII-spectrin ( Sptan1 ) knockdown at low tension (low-dose blebbistatin 5 µM) showing streak-like defects similar to in vivo. Representative images of n = 3 biological replicates each. (g) Illustration of laser ablation in multilayered keratinocytes. (h) Laser ablation: Still images from live imaging of SiR-actin (F-actin)-labeled Ctr cells (48 h high Ca 2+ ) at indicated time points after 17 µm line ablation showing progressive elliptical cortical openings. (i) Quantification of the elliptical opening area (red line) i.e., recoil over time. Line: mean ± SD opening area of 13 ablations from N = 4 biological replicates. (j) Cortical laser ablation: Still images from live imaging of SiR-actin (F-actin)–labeled Ctr (as shown h) and αII-spectrin knockdown keratinocytes at the time point of analysis (60 s after a linear laser cut [yellow line]) after 48 h high Ca 2+ . The opening in the F-actin cortex is seen as black area around the yellow line. (k) Quantification of opening areas in the apical F-actin cortex upon linear laser ablation as shown in j. Lines represent means, dots represent single openings/cells pooled from n = 3 independent experiments/biological replicates. *P = 0.0215 with Kolmogorov–Smirnov. (l) Laser ablation (as described for j) of αII-spectrin knockdown keratinocytes treated with DMSO or low dose blebbistatin (5 µM) 1 h prior to ablation. (m) Quantification of opening areas in the apical F-actin cortex upon linear laser ablation as shown in l. Lines represent means; dots represent single openings/cells pooled from n = 3 independent experiments/biological replicates. ****P < 0.0001 with Kolmogorov–Smirnov. (n) Laser ablation (as described for j) of E-cadherin −/− keratinocytes after 48 h high Ca 2+ . (o) Quantification of opening areas in the apical F-actin cortex upon linear laser ablation as shown in N . Lines represent means; dots represent single openings/cells pooled from n = 3 independent experiments/biological replicates. ****P < 0.0001 with Kolmogorov–Smirnov. ROI, region of interest.

Article Snippet: At confluency, the medium was switched to a high Ca 2+ medium for 48 h. 2 h prior to imaging, the medium was changed to a high Ca 2+ medium containing 1 μM SiR-actin (#CY-SC001; Spirochrome).

Techniques: Immunofluorescence, Immunolabeling, Microscopy, Knockdown, In Vivo, Imaging, Labeling

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