Journal: Pharmacological Research

Article Title: Isoginkgetin protects against degeneration of ALS motor neurons via regulating the GSK-3β–TFEB signaling axis

doi: 10.1016/j.phrs.2026.108172

Figure Lengend Snippet: Lysosomal dysfunction in samples from patients with ALS and iPSC-derived motor neurons. (A) Western blotting was performed to detect the expression of lysosome-related proteins in the brain tissues (Middle frontal gyrus) from patients with ALS and healthy controls. The results revealed lysosomal dysfunction in the samples from patients with ALS. (B) Immunostaining of motor neurons derived from eight iPSC lines on day 10 in stage 5 demonstrating the differentiation of MAP2 + /HB9 + motor neurons. Cellular nuclei were counterstained with DAPI. Scale bars, 100 μm. (C) Schematic diagram of magnetic microbead sorting for motor neurons. As observed in the immunofluorescence and bright-field images, MAP2 + /HB9 + motor neurons displayed significant enrichment. Scale bars, 100 μm. (D) Western blotting analysis was performed to detect the expression of lysosome-related proteins in purified day-28 ALS motor neurons and healthy controls. The results indicated lysosomal dysfunction in ALS motor neurons. Health ctrl 1: GZF2; Health ctrl 2: 2–8–8–2; Health ctrl 3: UC12; Health ctrl 4: UC01; ALS 1: TDP-43 A315T; ALS 2: TDP-43 A382T; ALS 3: SOD1 G94A; ALS 4: SOD1 D90A. (E) Representative live-cell imaging images of 28-day-old motor neurons labeled with Syn ::EGFP and treated with LysoTracker Red dye. Scale bars, 10 μm. (F) Quantitative results of (E) showed a significant reduction in the number of functional lysosomes in ALS motor neurons; n = 20 motor neurons for each group. (G) Representative live-cell imaging images of 28-day-old motor neurons labeled with Syn ::EGFP and treated with DQ-BSA-Red dye. Scale bars, 10 μm. (H) Quantitative results of (G) showed a significant decrease in the lysosomal activity in ALS motor neurons; n = 20 motor neurons for each group. Data were presented as the mean ± standard deviation. Two-way ANOVA was followed by Tukey’s multiple comparison test.

Article Snippet: Two iPSC lines from patients with ALS, which were purchased from WiCell Research Institute Inc., harbored the following mutations: TDP-43 (PFIZi013-A, TARDBP A382T ) and SOD-1 (WC034i, SOD1 D90A ).

Techniques: Derivative Assay, Western Blot, Expressing, Immunostaining, Immunofluorescence, Purification, Live Cell Imaging, Labeling, Functional Assay, Activity Assay, Standard Deviation, Comparison

Journal: Pharmacological Research

Article Title: Isoginkgetin protects against degeneration of ALS motor neurons via regulating the GSK-3β–TFEB signaling axis

doi: 10.1016/j.phrs.2026.108172

Figure Lengend Snippet: ISO improves lysosomal function in ALS motor neurons via the GSK-3β–TFEB signaling axis. (A) Representative immunofluorescence images for detecting TFEB nuclear translocation induced by ISO (750 nM, 24 h) in 28-day-old ALS motor neurons. Scale bars, 20 μm. (B) Quantitative results of (A) showed that ISO induced TFEB nuclear translocation in four types of ALS motor neurons; n = 20 motor neurons for each group. (C) Western blotting was performed to detect the expression of lysosome-related proteins in purified day-28 ALS motor neurons treated with ISO (750 nM, 24 h). The results indicated that ISO inhibited GSK-3β activity and increased the expression of lysosome-related proteins. Sample 1: TDP-43 A315T; Sample 2: TDP-43 A382T; Sample 3: SOD1 G94A; and Sample 4: SOD1 D90A. (D) Representative live-cell imaging images of 28-day-old motor neurons labeled with Syn ::EGFP and treated with LysoTracker Red dye after ISO (750 nM, 24 h) treatment. Scale bars, 10 μm. (E) Quantitative results of (D) showed that ISO increased the number of functional lysosomes in ALS motor neurons; n = 20 motor neurons for each group. (F) Representative live-cell imaging images of 28-day-old motor neurons labeled with Syn ::EGFP and treated with DQ-BSA-Red dye after ISO (750 nM, 24 h) treatment. Scale bars, 10 μm. (G) Quantitative results of (F) showed that ISO enhanced the lysosomal activity in ALS motor neurons; n = 20 motor neurons for each group. (H) Representative images of neurite swelling in ALS motor neurons after 7-day ISO (750 nM) treatment detected by immunofluorescence (white arrow). Scale bars, 50 μm. (I) Quantitative results of (H) showed that ISO reduced the proportion of neurite bead-like swelling in ALS motor neurons; n = 20 images for each group. (J) Quantification results of the area of Syn ::EGFP-labeled ALS motor neurons from day 28 to day 35 showed a gradual loss of EGFP signals, which was attributed to the progressive death of ALS motor neurons; n = 20 images for each group. (K) Representative images of Syn ::EGFP-labeled ALS motor neurons, captured via live-cell imaging after 7-day ISO (750 nM, from day 28 to day 35) treatment. Scale bars, 200 μm. (L) Quantitative results of (K) showed that ISO increased the number of surviving Syn :: EGFP + ALS motor neurons; n = 20 images for each group. Data were presented as the mean ± standard deviation. Two-way ANOVA was followed by Tukey’s multiple comparison test.

Article Snippet: Two iPSC lines from patients with ALS, which were purchased from WiCell Research Institute Inc., harbored the following mutations: TDP-43 (PFIZi013-A, TARDBP A382T ) and SOD-1 (WC034i, SOD1 D90A ).

Techniques: Immunofluorescence, Translocation Assay, Western Blot, Expressing, Purification, Activity Assay, Live Cell Imaging, Labeling, Functional Assay, Standard Deviation, Comparison

Journal: International Dental Journal

Article Title: Obesity as a Determinant of Periodontal Therapy Outcomes: Insights on Oxidative and Endoplasmic Reticulum Stress Pathways

doi: 10.1016/j.identj.2026.109472

Figure Lengend Snippet: Oxidative stress markers in the study population, stratified by obesity presence, before and after non-surgical periodontal treatment. Total ROS (A and G), cytosolic superoxide (B and H) and mitochondrial superoxide (C and I) in leukocytes, and protein levels of SOD1 (D and J) in PBMCs normalized to the loading control actin with corresponding WB images (E), and serum total antioxidant capacity (F and K). Data are presented as box and whisker plots and were compared using two-factor analysis of variance (ANOVA) followed by post hoc tests. ** P < .01 when comparing baseline vs 12 wk. # P < .05 when comparing groups with vs without obesity. DCFH, diclorodihidrofluoresceína; dHE, dihydroethidium; MitoSOX, Mitochondrial Superoxide Indicator; ns, not significant; PBMCs, peripheral blood mononuclear cells; ROS, reactive oxygen species; SOD1, superoxide dismutase 1.

Article Snippet: Twenty-five micrograms of protein were separated by SDS-PAGE, transferred to nitrocellulose membranes, blocked, and incubated with the following specific antibodies: rabbit monoclonal anti-superoxide dismutase 1 (SOD1) (Cat# PA5-27240, RRID:AB_2544716), rabbit polyclonal anti-IRE1α (Cat# PA1-46027, RRID:AB_2262265), mouse monoclonal anti-CHOP (Cat# MA1-250, RRID:AB_2292611) from Thermo Fisher Scientific; rabbit polyclonal anti-GRP78 (Cat# ab21685, RRID:AB_2119834), mouse monoclonal anti-ATF6 (Cat# ab122897, RRID:AB_10899) from Abcam; rabbit polyclonal anti-eukaryotic initiation factor 2 phospho (p-eIF2α) (Innovative Research Cat# 44-728G, RRID:AB_15000); mouse monoclonal anti-actin (Cell Signaling Technology Cat# 3700, RRID:AB_2242334) or rabbit polyclonal anti-actin (Sigma-Aldrich Cat# A5060, RRID:AB_476738).

Techniques: Control, Whisker Assay