Journal: EMBO Molecular Medicine

Article Title: The HSP40 chaperone Ydj1 drives amyloid beta 42 toxicity

doi: 10.15252/emmm.202113952

Figure Lengend Snippet: A Pearson’s correlation coefficient and Manders’ co‐localization Coefficients of endoplasmatic reticulum protein KDEL and Abeta analyzed on confocal images of Kenyon cells in 15‐day‐old male fly brains immunostained with Abeta‐specific antibody (Abeta) 6E10 and KDEL‐specific antibody in Droj2 knockdown flies ( Droj2 +/− ) and corresponding isogenic w 1118 wild‐type flies ( Droj2 +/+ ) with expression of human Abeta42 (UAS‐A42). Dot plots show all data points along with the mean (bar) ± SD n = 10. Unpaired, two‐tailed t ‐test. B Representative confocal and gSTED deconvolved (decon) images of Kenyon cells in 15‐day‐old male fly brains immunostained with Abeta‐specific antibody (Abeta) 6E10 (magenta) and endoplasmic reticulum protein KDEL‐specific antibody (KDEL, green) of Droj2 knockdown flies ( Droj2 +/− ) and corresponding isogenic w 1118 wild‐type flies ( Droj2 +/+ ) expressing human Abeta42 (UAS‐A42). C Representative confocal and gSTED deconvolved (decon) microscopy of Kenyon cells in 15‐day‐old male fly brains immunostained with Abeta‐specific antibody (Abeta) 6E10 (magenta) and mitochondrial marker ATP5A‐specific antibody (ATP5A, green) of w 1118 wild‐type flies ( Droj2 +/+ ) expressing human Abeta42 (UAS‐A42). D–H Counts of mitochondria (D), mitochondria average size (E), mitochondria coverage area (F), Feret diameter (G), and min Feret diameter (H) of ATP5A‐stained mitochondria from fly brain gSTED deconvolved images representatively shown in Fig from 9 to 10 brains of w 1118 wild‐type ( Droj2 +/+ ) and knockdown ( Droj2 +/− ) male flies expressing human Abeta42 (UAS‐A42). Dot plots show all data points along with the mean (bar) ± SD n = 9–10. Unpaired, two‐tailed t ‐test or Mann–Whitney test.

Article Snippet: The following primary antibodies were used: mouse monoclonal IgG1 κ anti‐Aß antibody (clone 6E10; Biolegend, 803001; 1:200 for confocal, 1:400 for gSTED), mouse IgG2b anti‐ATP5A (clone 15H4C4, Abcam, ab14748, 1:100 for gSTED), and mouse IgG2a anti‐KDEL (clone 10C3, Enzo Lifesciences, ADI‐SPA‐827, 1:100 for gSTED).

Techniques: Expressing, Two Tailed Test, Microscopy, Marker, Staining, MANN-WHITNEY

Journal: Nutrients

Article Title: Anti-Inflammatory Activity of Isomaltodextrin in a C57BL/6NCrl Mouse Model with Lipopolysaccharide-Induced Low-Grade Chronic Inflammation

doi: 10.3390/nu11112791

Figure Lengend Snippet: Effect of IMD on PPAR-γ and IRS-1expression in white adipose tissue of mice as determined by Western blot. Results are expressed as mean ± SEM for n = 4 samples per group. Differences in means were considered statistically significant for * p < 0.05 or ** p < 0.01.

Article Snippet: The membranes were washed 5 times, for 5 minutes each, and incubated with the secondary antibody diluted in 3% BSA in TBST (1:2000 ( v / v ) diluted anti-mouse for PPAR-γ and TLR-4, or 1:1000 ( v / v ) diluted anti-rabbit for IRS-1 (Promega, Madison, WI)) for one hour at room temperature with gentle shaking.

Techniques: Western Blot

Journal: Frontiers in Immunology

Article Title: Electroacupuncture alleviates pain by activating the MD2/TLR4/NF-κB pathway in the ST36 acupoint

doi: 10.3389/fimmu.2025.1626755

Figure Lengend Snippet: EA-induced analgesia is associated with MD2, TLR4, and p-p65/p65 in the ST36 acupoint and MD2 and TLR4 interactions. (A) The experimental protocol timeline is illustrated, with the duration of each intervention clearly indicated in the figure. (B) The location of the ST36 acupoint in mice (up) and the swelling of paw comparison before (right) and after (left) CFA injection. (C) PTWTs were detected on the D-3 to D-1, D1pre, D1post, D3, D5, and D7 after interventions (n = 8), ** P < 0.01 vs. the CON, ※※ P < 0.01 vs. the SHAM. (D) The expression levels of PKC protein in the spinal cord were determined by WB (n = 3), ** P < 0.01 vs. the CON, ** P < 0.01 vs. the SHAM. (E, F) The expression levels of CX3CL1 (E) , IL-1β (F) in the spinal cord were determined by Elisa (n = 6), ** P < 0.01 vs. the CON, ** P < 0.01 vs. the SHAM. (G) Data sourced from https://cn.string-db.org/ . (H) Co-IP assay with TLR4 antibody (or IgG) in the ST36 acupoint, followed by WB analysis of MD2 and TLR4. (I–K) The expression levels of MD2 (I) , TLR4 (J) , and p-p65/p65 (K) were determined using WB analysis in the ST36 acupoint on D7 (n = 3), * P < 0.05 vs. the CON, ** P < 0.01 vs. the CON, ※※ P < 0.01 vs. the SHAM.

Article Snippet: Tissue lysate (1 mg protein) was incubated with either 2 μg of mouse monoclonal anti-TLR4 antibody or 2 μg of mouse IgG isotype control antibody (5415, Cell Signaling Technology, USA) for 1 hour at room temperature.

Techniques: Comparison, Injection, Expressing, Enzyme-linked Immunosorbent Assay, Co-Immunoprecipitation Assay

Journal: Frontiers in Immunology

Article Title: Electroacupuncture alleviates pain by activating the MD2/TLR4/NF-κB pathway in the ST36 acupoint

doi: 10.3389/fimmu.2025.1626755

Figure Lengend Snippet: The effects of modulating MD2 in the ST36 acupoint on EA-induced analgesia and on the protein levels of MD2, TLR4, and p-p65/p65. (A) Lentivirus injection, behavioral testing, and acupoint stimulation experimental schedule. (B) Detection of lentivirus transduction in the ST36 acupoint by IF assay after MD2 knockdown lentivirus injection (n=3). (C) IF assay detection of lentivirus transduction in ST36 acupoint after overexpression lentivirus injection (n=3). (D) The transfection efficiency of the control lentivirus (FR-376) in ST36 acupoint (n=3). (E) The transfection efficiency of the control lentivirus (FV-115) in ST36 acupoint (n=3). (F) The expression of MD2 after injection of three different target knockdown lentiviruses were detected by WB (n=4). (G) The expression of MD2 after injection of overexpression was detected by WB (n=3). (H) The PTWTs were detected on D-3 to D-1, D1pre, D1post to D7 (every other day) (n = 6), ** P < 0.01 vs. the AIA+LVcon, # P < 0.05 vs. the AIA+LVcon+EA, ## P < 0.01 vs. the AIA+LVcon+EA. (I–K) The expression level of MD2 (I) , TLR4 (J) , and p-p65/p65 (K) (n = 3), ** P < 0.01 vs. the AIA+LVcon, # P < 0.05 vs. the AIA+LVcon+EA, ## P < 0.01 vs. the AIA+LVcon+EA. (L) PTWTs were detected on D-3 to D-1, D1pre, D1post to D7 (every other day) (n = 6), * P < 0.05 vs. the AIA+OEcon, ** P < 0.01 vs. the AIA+OEcon. (M–O) The expression level of MD2 (M) , TLR4 (N) , and p-p65/p65 (O) (n = 3), ** P < 0.01 vs. the AIA+OEcon.

Article Snippet: Tissue lysate (1 mg protein) was incubated with either 2 μg of mouse monoclonal anti-TLR4 antibody or 2 μg of mouse IgG isotype control antibody (5415, Cell Signaling Technology, USA) for 1 hour at room temperature.

Techniques: Injection, Transduction, Knockdown, Over Expression, Transfection, Control, Expressing

Journal: Frontiers in Immunology

Article Title: Electroacupuncture alleviates pain by activating the MD2/TLR4/NF-κB pathway in the ST36 acupoint

doi: 10.3389/fimmu.2025.1626755

Figure Lengend Snippet: EA treatment can increase the co-expression of MD2/TLR4, TLR4-Vimentin, TLR4-Tryptase, as well as TLR4-F4/80 at the ST36 acupoint. (A) Double-labeling immunofluorescence of TLR4 (green), MD2 (red) and DAPI (blue), scale bar = 20 µm. (B) Double-labeling immunofluorescence of TLR4 (green), fibroblasts (red) and DAPI (blue), scale bar = 20 µm. (C) Double-labeling immunofluorescence of TLR4 (green) and Tryptase (red), scale bar = 20µm. (D) Double-labeling immunofluorescence of TLR4 (green) and F4/80 (red), DAPI (blue), scale bar = 20µm. (E) Immunofluorescence quantitative analysis of MD2/TLR4 (n = 3), * P < 0.05 vs. the CON, ## P < 0.01 vs. the AIA. (F) Immunofluorescence quantitative analysis of TLR4-Vimentin (n = 3), ** P < 0.01 vs. the CON, ## P < 0.01 vs. the AIA. (G) Immunofluorescence quantitative analysis of TLR4-Tryptase (n = 3), # P < 0.05 vs. the AIA. (H) Immunofluorescence quantitative analysis of TLR4-F4/80 (n = 3).

Article Snippet: Tissue lysate (1 mg protein) was incubated with either 2 μg of mouse monoclonal anti-TLR4 antibody or 2 μg of mouse IgG isotype control antibody (5415, Cell Signaling Technology, USA) for 1 hour at room temperature.

Techniques: Expressing, Labeling, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: Electroacupuncture alleviates pain by activating the MD2/TLR4/NF-κB pathway in the ST36 acupoint

doi: 10.3389/fimmu.2025.1626755

Figure Lengend Snippet: The pattern of the effect of EA treatment on MD2, TLR4 in ST36 acupoint. (A) Behavioral testing, acupoint stimulation and molecular detection experimental schedule. (B–D) PTWTs were detected on D1post (B) (n=8), D3 (C) (n=6), and D5 (D) (n=6), ** P < 0.01 vs. the CON, ※ P < 0.05 vs. the SHAM, ※※ P < 0.01 vs. the SHAM. (E–H) The expression level of MD2 in the ST36 acupoint was determined by WB on D1post (E) , D3 (F) and D5 (G) (n = 3); the TLR4 was determined on the D5 (H) (n = 3), * P < 0.05 vs. the CON, ** P < 0.01 vs. the CON, # P < 0.05 vs. the AIA. ※ P < 0.05 vs. the SHAM.

Article Snippet: Tissue lysate (1 mg protein) was incubated with either 2 μg of mouse monoclonal anti-TLR4 antibody or 2 μg of mouse IgG isotype control antibody (5415, Cell Signaling Technology, USA) for 1 hour at room temperature.

Techniques: Expressing

Journal: BMC Pregnancy and Childbirth

Article Title: Maternal undernourishment impairs murine placental development during pregnancy

doi: 10.1186/s12884-025-07951-z

Figure Lengend Snippet: Increased ROS and unchanged TLR4 abundance in MUN placentas. ( A ) Representative images and ( B ) quantification of CellROX staining for ROS ( n = 4 dams, * p ≤ 0.05 vs. Control, # p ≤ 0.05 vs. MUN) in the labyrinth zone of murine placentas. ( C ) Representative western blot images and ( D ) quantification of TLR4 abundance in placental samples ( n = 4 dams) normalized to GAPDH. Western blot samples were derived from the same experiment, processed in parallel alongside controls, and cropped for clarity. Adjustments to the contrast and brightness of representative images were applied equally across the entire image and optimized for publication purposes only. Negative control images for placental immunohistochemistry can be found in Supplemental Figure . Abbreviations: ROS, reactive oxygen species; MUN, maternal undernourishment; Mito, MitoTEMPO; TAK, TAK-242. Placenta schematic created in BioRender. Camilliere, M. (2025) https://BioRender.com/b86b146

Article Snippet: Primary antibodies used included: rabbit anti-mouse Catalase (1:1,000) (Cell Signaling, Danvers, MA), rabbit anti-mouse SOD1 (1:1,000) (Cell Signaling), rabbit anti-mouse TLR4 (1:1,000) (Cell Signaling), rabbit anti-mouse CD31 (1:1,000) (Cell Signaling), rabbit anti-mouse E-Cadherin (1:1,000) (Cell Signaling), mouse anti-mouse VEGF (1:1000) (Invitrogen) and mouse anti-mouse GAPDH (1:5,000) (Millipore).

Techniques: Staining, Control, Western Blot, Derivative Assay, Negative Control, Immunohistochemistry