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anti lc3 i ii  (Proteintech)


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    Structured Review

    Proteintech anti lc3 i ii
    Anti Lc3 I Ii, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 2126 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lc3+i+ii/LC3+Antibody/pm42767428-127-6-10
    Average 96 stars, based on 2126 article reviews
    anti lc3 i ii - by Bioz Stars, 2026-11
    96/100 stars

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    Related Articles

    Blocking Assay:

    Article Title: Identification of ferroptosis-related genes as diagnostic biomarkers and therapeutic targets in sorafenib-resistant HCC via machine learning and experimental validation.
    Article Snippet: Identification of ferroptosis-related genes as diagnostic biomarkers and therapeutic targets in sorafenib-resistant HCC via machine learning and experimental validation Jinrui Zhang 1, Wenhan Geng 1, Haojin Sun 1, Jian Chen 1, Yumei Fan *, Ke Tan * Ministry of Education Key Laboratory of Molecular and Cellular Biology; Hebei Research Center of the Basic Discipline of Cell Biology, Hebei Collaborative Innovation Center for Eco-Environment, Hebei Province Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology, College of Life Sciences, Hebei Normal University, Shijiazhuang, China A R T I C L E I N F O Editor: Jean-Pierre GORVEL Keywords: sorafenib resistant ferroptosis diagnosis machine learning algorithms A B S T R A C T Liver cancer, particularly hepatocellular carcinoma (HCC), remains a significant health burden with poor survival rates due to late diagnosis and limited treatment options.. Sorafenib, a multi-kinase inhibitor, is the standard first-line therapy for advanced HCC, but long-term use often leads to drug resistance.. Ferroptosis, an irondependent form of regulated cell death, has emerged as a crucial target in cancer therapy, showing potential to enhance treatment efficacy and overcome drug resistance.

    Incubation:

    Article Title: Identification of ferroptosis-related genes as diagnostic biomarkers and therapeutic targets in sorafenib-resistant HCC via machine learning and experimental validation.
    Article Snippet: Identification of ferroptosis-related genes as diagnostic biomarkers and therapeutic targets in sorafenib-resistant HCC via machine learning and experimental validation Jinrui Zhang 1, Wenhan Geng 1, Haojin Sun 1, Jian Chen 1, Yumei Fan *, Ke Tan * Ministry of Education Key Laboratory of Molecular and Cellular Biology; Hebei Research Center of the Basic Discipline of Cell Biology, Hebei Collaborative Innovation Center for Eco-Environment, Hebei Province Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology, College of Life Sciences, Hebei Normal University, Shijiazhuang, China A R T I C L E I N F O Editor: Jean-Pierre GORVEL Keywords: sorafenib resistant ferroptosis diagnosis machine learning algorithms A B S T R A C T Liver cancer, particularly hepatocellular carcinoma (HCC), remains a significant health burden with poor survival rates due to late diagnosis and limited treatment options.. Sorafenib, a multi-kinase inhibitor, is the standard first-line therapy for advanced HCC, but long-term use often leads to drug resistance.. Ferroptosis, an irondependent form of regulated cell death, has emerged as a crucial target in cancer therapy, showing potential to enhance treatment efficacy and overcome drug resistance.

    Article Title: Novel Isolongifolenone-Based Caprolactam Derivatives as Potential Anticancer Agents via the p53/mTOR/Autophagy Pathway.
    Article Snippet: .. The membranes were then blocked with BSA for 2 h at room temperature and incubated with primary antibodies against p-mTOR (ImmunoWay Biotechnology, Beijing, China, YP1444, 1:5000), P62 (Proteintech Group, Inc., Wuhan, China, 60268-1-IG, 1:5000), LC3-I/II (Proteintech Group, Inc., Wuhan, China, 10253-2-AP, 1:5000) and p-AMPK, AMPK, Bcl-2, Bax, Cleaved-caspase 7 (ImmunoWay Biotechnology, Beijing, China, YP0251, 1:5000) overnight at 4 ◦C and with secondary antibodies (AP-conjugated goat anti-rabbit or anti-mouse, 1:10,000) for 2 h. Densitometry was performed in ImageJ 2 within the linear range. ..

    Article Title: Novel Isolongifolenone-Based Caprolactam Derivatives as Potential Anticancer Agents via the p53/mTOR/Autophagy Pathway
    Article Snippet: .. The membranes were then blocked with BSA for 2 h at room temperature and incubated with primary antibodies against p-mTOR (ImmunoWay Biotechnology, Beijing, China, YP1444, 1:5000), P62 (Proteintech Group, Inc., Wuhan, China, 60268-1-IG, 1:5000), LC3-I/II (Proteintech Group, Inc., Wuhan, China, 10253-2-AP, 1:5000) and p-AMPK, AMPK, Bcl-2, Bax, Cleaved-caspase 7 (ImmunoWay Biotechnology, Beijing, China, YP0251, 1:5000) overnight at 4 °C and with secondary antibodies (AP-conjugated goat anti-rabbit or anti-mouse, 1:10,000) for 2 h. Densitometry was performed in ImageJ 2 within the linear range. ..

    Article Title: O-GlcNAcylation of YAP1 promotes lung transplant ischemia-reperfusion injury via binding to HIF1α transcription factor and activating autophagy and mitophagy
    Article Snippet: .. Afterward, the membrane underwent treatment with 5% skim milk powder at room temperature for 1 h. It was then incubated overnight at 4 °C with primary antibodies against YAP1 (Cat No. 13584-1-AP, 1:1500), p-YAP1 (Cat No. 80694-2-RR, 1:2000), HIF-1α (Cat No. 66730-1-Ig, 1:1000), CYR61 (No. 26689-1-AP, 1:1500), AREG (Cat No. 16036-1-AP, 1:2300), BIRC5 (Cat No. 10508-1-AP, 1:1200), FUNDC1 (Cat No. 28519-1-AP, 1:1500), PINK1 (Cat No. 23274-1-AP, 1:2000), TBK1 (Cat No. 28397-1-AP, 1:1500), HMGB1 (Cat No. 10829-1-AP, 1:1000), DAPK (Cat No. 25136-1-AP, 1:1000), LC3-I/II (Cat No. 14600-1-AP, 1:3000), OGT (Cat No. 11576-2-AP, 1:2000) and O-GlcNAc (Cat No. 65292-1-Ig, 1:3000) (all obtained from Proteintech, Wuhan, China). .. After the incubation period, the membrane underwent retesting using a secondary antibody IgG (Santa Cruz Biotechnology, Inc, Santa Cruz, CA) labeled with horseradish peroxidase.

    Article Title: O-GlcNAcylation of YAP1 promotes lung transplant ischemia-reperfusion injury via binding to HIF1α transcription factor and activating autophagy and mitophagy.
    Article Snippet: .. It was then incubated overnight at 4 °C with primary antibodies against YAP1 (Cat No. 13584-1-AP, 1:1500), p-YAP1 (Cat No. 80694-2-RR, 1:2000), HIF-1α (Cat No. 66730-1-Ig, 1:1000), CYR61 (No. 26689-1-AP, 1:1500), AREG (Cat No. 16036-1-AP, 1:2300), BIRC5 (Cat No. 10508-1-AP, AR TI CL E IN P RE SS 1:1200), FUNDC1 (Cat No. 28519-1-AP, 1:1500), PINK1 (Cat No. 23274-1-AP, 1:2000), TBK1 (Cat No. 28397-1-AP, 1:1500), HMGB1 (Cat No. 10829-1-AP, 1:1000), DAPK (Cat No. 25136- 1-AP, 1:1000), LC3-I/II (Cat No. 14600-1-AP, 1:3000), OGT (Cat No. 11576-2-AP, 1:2000) and O-GlcNAc (Cat No. 65292-1-Ig, 1:3000) (all obtained from Proteintech, Wuhan, China). .. After the incubation period, the membrane underwent retesting using a secondary antibody IgG (Santa Cruz Biotechnology, Inc, Santa Cruz, CA) labeled with horseradish peroxidase.

    other:


    Western Blot:

    Article Title: Overexpression of USP35 enhances the protective effect of hUC-MSCs and their extracellular vesicles in oxygen-glucose deprivation/reperfusion-induced SH-SY5Y cells via stabilizing FUNDC1
    Article Snippet: The band intensity was quantified by ImageJ software (NIH) and normalized to the housekeeping gene that served as internal control. .. The antibodies for Western Blot and their dilution are listed as follows: USP35 (Proteintech, Cat#: 24559-1-AP, 1:1000), FUNDC1 (Proteintech, Cat#: 28519-1-AP, 1:2000), Cyt C (Proteintech, Cat#: 10993-1-AP, 1:1000), BAX (Proteintech, Cat#: 50599-2-Ig, 1:1000), BCL2 (Proteintech, Cat#: 12789-1-AP, 1:2000), total and cleaved Caspase 3 (Proteintech, Cat#: 19677-1-AP, 1:1000), total and cleaved Caspase 9 (Proteintech, Cat#: 10380-1-AP, 1:1000), Beclin-1 (Proteintech, Cat#: 11306-1-AP, 1:1000), p62 (Proteintech, Cat#: 18420-1-AP, 1:5000), LC3 I/II (Proteintech, Cat#: 14600-1-AP, 1:1000), Flag (Proteintech, Cat#: 20543-1-AP, 1:20000), HA (Proteintech, Cat#: 51064-2-AP, 1:5000), His (Proteintech, Cat#: 10001-0-AP, 1:1000), β-actin (Proteintech, Cat#: 20536-1-AP, 1:1000), Luciferase (Proteintech, Cat#: 27986-1-AP, 1:1000), CD9 (Proteintech, Cat#: 20597-1-AP, 1:1000), CD81 (Proteintech, Cat#: 27855-1-AP, 1:1000), CD63 (Proteintech, Cat#: 25682-1-AP, 1:1000), horseradish peroxidase-conjugated goat anti-rabbit IgG (Proteintech, Cat#: SA00001-2, 1:2000). ..

    Luciferase:

    Article Title: Overexpression of USP35 enhances the protective effect of hUC-MSCs and their extracellular vesicles in oxygen-glucose deprivation/reperfusion-induced SH-SY5Y cells via stabilizing FUNDC1
    Article Snippet: The band intensity was quantified by ImageJ software (NIH) and normalized to the housekeeping gene that served as internal control. .. The antibodies for Western Blot and their dilution are listed as follows: USP35 (Proteintech, Cat#: 24559-1-AP, 1:1000), FUNDC1 (Proteintech, Cat#: 28519-1-AP, 1:2000), Cyt C (Proteintech, Cat#: 10993-1-AP, 1:1000), BAX (Proteintech, Cat#: 50599-2-Ig, 1:1000), BCL2 (Proteintech, Cat#: 12789-1-AP, 1:2000), total and cleaved Caspase 3 (Proteintech, Cat#: 19677-1-AP, 1:1000), total and cleaved Caspase 9 (Proteintech, Cat#: 10380-1-AP, 1:1000), Beclin-1 (Proteintech, Cat#: 11306-1-AP, 1:1000), p62 (Proteintech, Cat#: 18420-1-AP, 1:5000), LC3 I/II (Proteintech, Cat#: 14600-1-AP, 1:1000), Flag (Proteintech, Cat#: 20543-1-AP, 1:20000), HA (Proteintech, Cat#: 51064-2-AP, 1:5000), His (Proteintech, Cat#: 10001-0-AP, 1:1000), β-actin (Proteintech, Cat#: 20536-1-AP, 1:1000), Luciferase (Proteintech, Cat#: 27986-1-AP, 1:1000), CD9 (Proteintech, Cat#: 20597-1-AP, 1:1000), CD81 (Proteintech, Cat#: 27855-1-AP, 1:1000), CD63 (Proteintech, Cat#: 25682-1-AP, 1:1000), horseradish peroxidase-conjugated goat anti-rabbit IgG (Proteintech, Cat#: SA00001-2, 1:2000). ..

    Membrane:

    Article Title: O-GlcNAcylation of YAP1 promotes lung transplant ischemia-reperfusion injury via binding to HIF1α transcription factor and activating autophagy and mitophagy
    Article Snippet: .. Afterward, the membrane underwent treatment with 5% skim milk powder at room temperature for 1 h. It was then incubated overnight at 4 °C with primary antibodies against YAP1 (Cat No. 13584-1-AP, 1:1500), p-YAP1 (Cat No. 80694-2-RR, 1:2000), HIF-1α (Cat No. 66730-1-Ig, 1:1000), CYR61 (No. 26689-1-AP, 1:1500), AREG (Cat No. 16036-1-AP, 1:2300), BIRC5 (Cat No. 10508-1-AP, 1:1200), FUNDC1 (Cat No. 28519-1-AP, 1:1500), PINK1 (Cat No. 23274-1-AP, 1:2000), TBK1 (Cat No. 28397-1-AP, 1:1500), HMGB1 (Cat No. 10829-1-AP, 1:1000), DAPK (Cat No. 25136-1-AP, 1:1000), LC3-I/II (Cat No. 14600-1-AP, 1:3000), OGT (Cat No. 11576-2-AP, 1:2000) and O-GlcNAc (Cat No. 65292-1-Ig, 1:3000) (all obtained from Proteintech, Wuhan, China). .. After the incubation period, the membrane underwent retesting using a secondary antibody IgG (Santa Cruz Biotechnology, Inc, Santa Cruz, CA) labeled with horseradish peroxidase.



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    Mice were orally administered αKG prior to establishment of a S. aureus -induced mastitis model. Mammary gland tissues were subsequently collected for analysis (n = 7 per group). A: Mitochondrial superoxide levels in frozen mammary gland tissue sections were assessed using MitoSOX Red staining, with green fluorescence indicating ROS. B: ATP levels in mammary gland tissues from Control, S. au, and αKG + S. au . C-D: OCR of mammary gland tissue cells in the three groups. E: Expression of <t>LC3,</t> ATG5, and p62 in protein lysates of mammary tissues from the three groups. G-H: RAW264.7 cells cultured with αKG were treated with oligomycin A and stimulated with S. aureus . Inflammatory cytokine levels and NF-κB signaling pathway activity of Control, S. au, αKG + S. au, αKG + S. au +oligomycin A were measured. I-P: qPCR analysis of M2 and M1 marker gene mRNA expression in four experimental groups of RAW264.7 cells. The data are presented as mean ± SD. One-way analysis of variance was performed for statistical analysis. * P < 0.05, ** P < 0.01 and *** P < 0.001 indicate significance from each group. ns, no significance.
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    Image Search Results


    CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, and enhances autophagy flux in PC12 cells. (A) The Western blot (WB) analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in PC12 cells from the CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (B) The Quantification of p-mTOR/t-mTOR protein levels. (C) The Quantification of p-AMPK/t-AMPK protein levels. (D) The WB analysis of LC3-I and LC3-II expression. (E) The Quantification of the LC3-II/LC3-I ratio. (F) The Comparison of LC3-II/LC3-I levels in the Baf-A1, CLE, and Baf-A1+CLE groups, assessing autophagic flux. (G) The WB analysis of p62 expression. (H) The Quantification of p62 protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.

    Journal: Frontiers in Aging Neuroscience

    Article Title: Clemastine alleviates Parkinson’s disease pathology by activating the AMPK/mTOR pathway and enhancing autophagic flux

    doi: 10.3389/fnagi.2026.1915475

    Figure Lengend Snippet: CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, and enhances autophagy flux in PC12 cells. (A) The Western blot (WB) analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in PC12 cells from the CTRL, MPP + , CLE, and MPP + +CLE groups. GAPDH is shown as a loading control. (B) The Quantification of p-mTOR/t-mTOR protein levels. (C) The Quantification of p-AMPK/t-AMPK protein levels. (D) The WB analysis of LC3-I and LC3-II expression. (E) The Quantification of the LC3-II/LC3-I ratio. (F) The Comparison of LC3-II/LC3-I levels in the Baf-A1, CLE, and Baf-A1+CLE groups, assessing autophagic flux. (G) The WB analysis of p62 expression. (H) The Quantification of p62 protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.

    Article Snippet: Proteins were transferred to PVDF membranes at 30 V for 90 min. Membranes were blocked with 5% non-fat milk in PBST for 1 h and incubated with primary antibodies for 2 h at room temperature: 1:50,000 mouse GAPDH (60004-1-Ig, Proteintech), 1:2,000 mouse p-mTOR (Ser2448) (67778-1-Ig, Proteintech), 1:5,000 mouse t-mTOR (66888-1-Ig, Proteintech), 1:1000 rabbit p-AMPK (Thr183/172) (ab133448, Abcam), 1:1000 mouse p62 (66184-1-Ig, Proteintech), 1:1,000 rabbit LC3-I/II (orb411847, Biorbyt), 1:5,000 mouse P62 (66184-1-Ig, Proteintech), 1:500 mouse α-syn (66412-1-Ig, Proteintech).

    Techniques: Phospho-proteomics, Western Blot, Analysis, Control, Expressing, Comparison

    CLE improves autophagy-related organelles in PC12 cells. (A) Immunofluorescence staining for LC3 (red), LAMP1 (green), and DAPI (blue) in PC12 cells from CTRL, MPP + , CLE, and MPP + +CLE groups. Lysosomes are defined as LAMP1 + LC3 – puncta; autophagosomes as LAMP1 + LC3 + puncta; autolysosomes as LAMP1 + LC3 + puncta. Scale bar: 5 μm. (B) The relative number of lysosomal puncta. (C) The relative number of autophagosome puncta. (D) The relative number of autolysosome puncta. Data are presented as mean ± SEM (N = 21 per group). Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; ANOVA, analysis of variance; SEM, standard error of the mean. n = 21 images per group (7 images from 3 independent experiments).

    Journal: Frontiers in Aging Neuroscience

    Article Title: Clemastine alleviates Parkinson’s disease pathology by activating the AMPK/mTOR pathway and enhancing autophagic flux

    doi: 10.3389/fnagi.2026.1915475

    Figure Lengend Snippet: CLE improves autophagy-related organelles in PC12 cells. (A) Immunofluorescence staining for LC3 (red), LAMP1 (green), and DAPI (blue) in PC12 cells from CTRL, MPP + , CLE, and MPP + +CLE groups. Lysosomes are defined as LAMP1 + LC3 – puncta; autophagosomes as LAMP1 + LC3 + puncta; autolysosomes as LAMP1 + LC3 + puncta. Scale bar: 5 μm. (B) The relative number of lysosomal puncta. (C) The relative number of autophagosome puncta. (D) The relative number of autolysosome puncta. Data are presented as mean ± SEM (N = 21 per group). Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. CLE, clemastine; CTRL, control; ANOVA, analysis of variance; SEM, standard error of the mean. n = 21 images per group (7 images from 3 independent experiments).

    Article Snippet: Proteins were transferred to PVDF membranes at 30 V for 90 min. Membranes were blocked with 5% non-fat milk in PBST for 1 h and incubated with primary antibodies for 2 h at room temperature: 1:50,000 mouse GAPDH (60004-1-Ig, Proteintech), 1:2,000 mouse p-mTOR (Ser2448) (67778-1-Ig, Proteintech), 1:5,000 mouse t-mTOR (66888-1-Ig, Proteintech), 1:1000 rabbit p-AMPK (Thr183/172) (ab133448, Abcam), 1:1000 mouse p62 (66184-1-Ig, Proteintech), 1:1,000 rabbit LC3-I/II (orb411847, Biorbyt), 1:5,000 mouse P62 (66184-1-Ig, Proteintech), 1:500 mouse α-syn (66412-1-Ig, Proteintech).

    Techniques: Immunofluorescence, Staining, Comparison, Control, Analysis

    CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, enhances autophagy, and reduces α-synuclein levels in the mouse midbrain. (A) The WB analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in the midbrain of mice from CTRL, MPTP, and MPTP+CLE groups. GAPDH is shown as a loading control. (B) The quantification of p-mTOR/t-mTOR levels. (C) The quantification of p-AMPK/t-AMPK levels. (D–F) The WB analysis of p62, LC3-I/II, and α-syn protein levels. (G–I) The quantification of p62, LC3-II/LC3-I ratio, and α-syn protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001. CLE, clemastine; α-syn, α-synuclein; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.

    Journal: Frontiers in Aging Neuroscience

    Article Title: Clemastine alleviates Parkinson’s disease pathology by activating the AMPK/mTOR pathway and enhancing autophagic flux

    doi: 10.3389/fnagi.2026.1915475

    Figure Lengend Snippet: CLE increases AMPK phosphorylation, decreases mTOR phosphorylation at Ser2448, enhances autophagy, and reduces α-synuclein levels in the mouse midbrain. (A) The WB analysis of p-AMPK (Thr172), t-AMPK, p-mTOR (Ser2448), and t-mTOR protein levels in the midbrain of mice from CTRL, MPTP, and MPTP+CLE groups. GAPDH is shown as a loading control. (B) The quantification of p-mTOR/t-mTOR levels. (C) The quantification of p-AMPK/t-AMPK levels. (D–F) The WB analysis of p62, LC3-I/II, and α-syn protein levels. (G–I) The quantification of p62, LC3-II/LC3-I ratio, and α-syn protein levels. Data are presented as mean ± SEM. Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001. CLE, clemastine; α-syn, α-synuclein; CTRL, control; WB, western blot; ANOVA, analysis of variance; SEM, standard error of the mean. n = 5 independent experiments.

    Article Snippet: Proteins were transferred to PVDF membranes at 30 V for 90 min. Membranes were blocked with 5% non-fat milk in PBST for 1 h and incubated with primary antibodies for 2 h at room temperature: 1:50,000 mouse GAPDH (60004-1-Ig, Proteintech), 1:2,000 mouse p-mTOR (Ser2448) (67778-1-Ig, Proteintech), 1:5,000 mouse t-mTOR (66888-1-Ig, Proteintech), 1:1000 rabbit p-AMPK (Thr183/172) (ab133448, Abcam), 1:1000 mouse p62 (66184-1-Ig, Proteintech), 1:1,000 rabbit LC3-I/II (orb411847, Biorbyt), 1:5,000 mouse P62 (66184-1-Ig, Proteintech), 1:500 mouse α-syn (66412-1-Ig, Proteintech).

    Techniques: Phospho-proteomics, Analysis, Control, Comparison, Western Blot

    CLE increases LC3 and reduces phosphorylated α-synuclein in dopaminergic neurons (DaNs) of MPTP-treated mice. (A) Immunofluorescence staining for TH (red, marker for DaNs), LC3 (green), and DAPI (blue) in the substantia nigra (SN) of mice from CTRL, MPTP, and MPTP+CLE groups. Scale bar: 2 μm. (B) The quantification of the relative number of LC3 puncta in DaNs. (C) Immunofluorescence staining for TH (red), p-α-syn (Ser129, green), and DAPI (blue) in the SN. Scale bar: 5 μm. (D) The quantification of the relative fluorescence intensity of p-α-syn (Ser129) in DaNs. Data are presented as mean ± SEM (N = 21 per group). Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001. CLE, clemastine; DaNs, dopaminergic neurons; p-α-syn, phosphorylated α-synuclein (Ser129); SN, substantia nigra; CTRL, control; ANOVA, analysis of variance; SEM, standard error of the mean. n = 21 images per group (7 images from 3 mice per group).

    Journal: Frontiers in Aging Neuroscience

    Article Title: Clemastine alleviates Parkinson’s disease pathology by activating the AMPK/mTOR pathway and enhancing autophagic flux

    doi: 10.3389/fnagi.2026.1915475

    Figure Lengend Snippet: CLE increases LC3 and reduces phosphorylated α-synuclein in dopaminergic neurons (DaNs) of MPTP-treated mice. (A) Immunofluorescence staining for TH (red, marker for DaNs), LC3 (green), and DAPI (blue) in the substantia nigra (SN) of mice from CTRL, MPTP, and MPTP+CLE groups. Scale bar: 2 μm. (B) The quantification of the relative number of LC3 puncta in DaNs. (C) Immunofluorescence staining for TH (red), p-α-syn (Ser129, green), and DAPI (blue) in the SN. Scale bar: 5 μm. (D) The quantification of the relative fluorescence intensity of p-α-syn (Ser129) in DaNs. Data are presented as mean ± SEM (N = 21 per group). Tukey’s multiple comparison test was performed after one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001. CLE, clemastine; DaNs, dopaminergic neurons; p-α-syn, phosphorylated α-synuclein (Ser129); SN, substantia nigra; CTRL, control; ANOVA, analysis of variance; SEM, standard error of the mean. n = 21 images per group (7 images from 3 mice per group).

    Article Snippet: Proteins were transferred to PVDF membranes at 30 V for 90 min. Membranes were blocked with 5% non-fat milk in PBST for 1 h and incubated with primary antibodies for 2 h at room temperature: 1:50,000 mouse GAPDH (60004-1-Ig, Proteintech), 1:2,000 mouse p-mTOR (Ser2448) (67778-1-Ig, Proteintech), 1:5,000 mouse t-mTOR (66888-1-Ig, Proteintech), 1:1000 rabbit p-AMPK (Thr183/172) (ab133448, Abcam), 1:1000 mouse p62 (66184-1-Ig, Proteintech), 1:1,000 rabbit LC3-I/II (orb411847, Biorbyt), 1:5,000 mouse P62 (66184-1-Ig, Proteintech), 1:500 mouse α-syn (66412-1-Ig, Proteintech).

    Techniques: Immunofluorescence, Staining, Marker, Fluorescence, Comparison, Control, Analysis

    Mice were orally administered αKG prior to establishment of a S. aureus -induced mastitis model. Mammary gland tissues were subsequently collected for analysis (n = 7 per group). A: Mitochondrial superoxide levels in frozen mammary gland tissue sections were assessed using MitoSOX Red staining, with green fluorescence indicating ROS. B: ATP levels in mammary gland tissues from Control, S. au, and αKG + S. au . C-D: OCR of mammary gland tissue cells in the three groups. E: Expression of LC3, ATG5, and p62 in protein lysates of mammary tissues from the three groups. G-H: RAW264.7 cells cultured with αKG were treated with oligomycin A and stimulated with S. aureus . Inflammatory cytokine levels and NF-κB signaling pathway activity of Control, S. au, αKG + S. au, αKG + S. au +oligomycin A were measured. I-P: qPCR analysis of M2 and M1 marker gene mRNA expression in four experimental groups of RAW264.7 cells. The data are presented as mean ± SD. One-way analysis of variance was performed for statistical analysis. * P < 0.05, ** P < 0.01 and *** P < 0.001 indicate significance from each group. ns, no significance.

    Journal: PLOS Pathogens

    Article Title: Glutamine alleviates Staphylococcus aureus -induced mastitis by modulating macrophage polarisation

    doi: 10.1371/journal.ppat.1014053

    Figure Lengend Snippet: Mice were orally administered αKG prior to establishment of a S. aureus -induced mastitis model. Mammary gland tissues were subsequently collected for analysis (n = 7 per group). A: Mitochondrial superoxide levels in frozen mammary gland tissue sections were assessed using MitoSOX Red staining, with green fluorescence indicating ROS. B: ATP levels in mammary gland tissues from Control, S. au, and αKG + S. au . C-D: OCR of mammary gland tissue cells in the three groups. E: Expression of LC3, ATG5, and p62 in protein lysates of mammary tissues from the three groups. G-H: RAW264.7 cells cultured with αKG were treated with oligomycin A and stimulated with S. aureus . Inflammatory cytokine levels and NF-κB signaling pathway activity of Control, S. au, αKG + S. au, αKG + S. au +oligomycin A were measured. I-P: qPCR analysis of M2 and M1 marker gene mRNA expression in four experimental groups of RAW264.7 cells. The data are presented as mean ± SD. One-way analysis of variance was performed for statistical analysis. * P < 0.05, ** P < 0.01 and *** P < 0.001 indicate significance from each group. ns, no significance.

    Article Snippet: The specific primary antibodies used in the experiment included p65 (#8242, Cell Signaling Technology, USA), p-p65 (#3033, Cell Signaling Technology, America), IκB (AF5002, Affinity Biosciences, Jiangsu, China), p-IκB (AF2002, Affinity Biosciences, Jiangsu, China), ZO-1 (AF5145, Affinity Biosciences, Jiangsu, China), Occludin (DF7504, Affinity Biosciences, Jiangsu, China), Claudin-3 (AF0129, Affinity Biosciences, Jiangsu, China), ATG5 (bsm-52596R, Bioss, Woburn, MA, USA), LC3-I/LC3-II (AF5402, Affinity Biosciences, Jiangsu, China), p62 (ab91526, Abcam plc, Cam bridge, UK), SOCS3 (bs-0580R, Bioss, Woburn, MA, USA), JAK1 (bs-1439R , Bioss, Woburn, MA, USA), p-JAK1 (bs-3238R, Bioss, Woburn, MA, USA), STAT3 (AF6294, Affinity Biosciences, Jiangsu, China), p-STAT3 (AF3293, Affinity Biosciences, Jiangsu, China) and β-actin (T0022, Affinity Biosciences, Jiangsu, China).

    Techniques: Staining, Fluorescence, Control, Expressing, Cell Culture, Activity Assay, Marker