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lc3 proteintech  (Proteintech)


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    Structured Review

    Proteintech lc3 proteintech
    Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for <t>LC3B</t> (green). Quantification of the mean fluorescence intensity of <t>LC3</t> is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001
    Lc3 Proteintech, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 2065 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/lc3+proteintech+14600+1+ap/LC3+Antibody/pmc12980994-96-58-59
    Average 96 stars, based on 2065 article reviews
    lc3 proteintech - by Bioz Stars, 2026-09
    96/100 stars

    Images

    1) Product Images from "S100A11 regulates microglial inflammatory response in neuropathic pain via H3K27ac-TFEB-mitochondrial autophagy axis"

    Article Title: S100A11 regulates microglial inflammatory response in neuropathic pain via H3K27ac-TFEB-mitochondrial autophagy axis

    Journal: The Journal of Headache and Pain

    doi: 10.1186/s10194-026-02328-9

    Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for LC3B (green). Quantification of the mean fluorescence intensity of LC3 is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001
    Figure Legend Snippet: Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for LC3B (green). Quantification of the mean fluorescence intensity of LC3 is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001

    Techniques Used: Knockdown, Western Blot, Labeling, Immunofluorescence, Staining, Fluorescence

    S100A11 knockdown alleviates microglial inflammation by inhibiting mitophagy through the TFEB/H3K27ac axis. A-B Western blot analysis of TFEB, TFEC, and TFE3 protein expression levels in the cytoplasm and nucleus of primary microglia, along with the corresponding detection and quantification plots ( D-F , for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). C Immunofluorescence staining of TFEB in primary microglia. G Western blot analysis of pink1, Parkin, P62, LC3II protein levels of primary microglia; H-K Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). M Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green). Quantification of the mean fluorescence intensity of LC3 is shown in panel L . and relative fluorescence intensity was quantified in panel L ( n = 3). N Western blot analysis of H4K16ac, H4K12ac, H3K27ac, H3K18ac, H3K9ac protein levels of microglia; O-S Relative levels normalized to histone H3 were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). T Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; U-X Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Z Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green), and relative fluorescence intensity was quantified in panel Y ( n = 3). Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001
    Figure Legend Snippet: S100A11 knockdown alleviates microglial inflammation by inhibiting mitophagy through the TFEB/H3K27ac axis. A-B Western blot analysis of TFEB, TFEC, and TFE3 protein expression levels in the cytoplasm and nucleus of primary microglia, along with the corresponding detection and quantification plots ( D-F , for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). C Immunofluorescence staining of TFEB in primary microglia. G Western blot analysis of pink1, Parkin, P62, LC3II protein levels of primary microglia; H-K Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). M Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green). Quantification of the mean fluorescence intensity of LC3 is shown in panel L . and relative fluorescence intensity was quantified in panel L ( n = 3). N Western blot analysis of H4K16ac, H4K12ac, H3K27ac, H3K18ac, H3K9ac protein levels of microglia; O-S Relative levels normalized to histone H3 were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). T Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; U-X Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Z Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green), and relative fluorescence intensity was quantified in panel Y ( n = 3). Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001

    Techniques Used: Knockdown, Western Blot, Expressing, Immunofluorescence, Staining, Labeling, Fluorescence

    Related Articles

    Ubiquitin Proteomics:

    Article Title: Targeting VCP with V8 suppresses glioblastoma development via formation of aggregates and disruption of mitophagy flux.
    Article Snippet: .. Primary antibodies included those against ACTB/βactin (ABclonal Technology, AC026), VCP(Proteintech, 10736–1-AP), XBP1S (Abcam, ab37152;1:2000), ATF4 (Proteintech, 10835–1-AP), P-EIF2α (Abclonal, AP0692), BiP/GRP78(Abclonal, A23453), IRE(Proteintech,27528– 1-AP), K48-linkage Specific Ubiquitin (Abclonal, A3606), PINK1(Proteintech, 23274–1-AP), PRKN (Abways, CY6641), P62(Proteintech, 18420–1-AP),BNIP3(Abclonal, A5683), LC3(Proteintech, 14600–1-AP), COXIV (HUABIO, ET1701-63), TAXIBP1(Proteintech, 14424–1-AP), TUBLIN (Abclonal, A17545), Ubiquitin (Proteintech, 10201–2-AP). .. Mitochondria protein were extracted from U87 cells using the Mitochondria Isolation Kit (Beyotime, C3601) according to the manufacturer’s instructions.

    Article Title: Targeting VCP with V8 suppresses glioblastoma development via formation of aggregates and disruption of mitophagy flux
    Article Snippet: .. Primary antibodies included those against ACTB/β-actin (ABclonal Technology, AC026), VCP(Proteintech, 10736–1-AP), XBP1S (Abcam, ab37152;1:2000), ATF4 (Proteintech, 10835–1-AP), P-EIF2α (Abclonal, AP0692), BiP/GRP78(Abclonal, A23453), IRE(Proteintech,27528–1-AP), K48-linkage Specific Ubiquitin (Abclonal, A3606), PINK1(Proteintech, 23274–1-AP), PRKN (Abways, CY6641), P62(Proteintech, 18420–1-AP),BNIP3(Abclonal, A5683), LC3(Proteintech, 14600–1-AP), COXIV (HUABIO, ET1701-63), TAXIBP1(Proteintech, 14424–1-AP), TUBLIN (Abclonal, A17545), Ubiquitin (Proteintech, 10201–2-AP). .. Mitochondria protein were extracted from U87 cells using the Mitochondria Isolation Kit (Beyotime, C3601) according to the manufacturer’s instructions.

    Article Title: The RNF8/OPTN/KDM6A axis controls macrophage polarization to maintain testicular microenvironment homeostasis.
    Article Snippet: 96), the semen supernatant samples of 5–6 healthy subjects and oligozoospermia patients were tested and statistically analyzed. qPCR Total RNA was extracted from testicular tissue and 293T cells using TRIZOL (Ambient, 95141801), and RNA concentration and purity were detected using nanodrop spectrophotometry (Thermo Scientific). cDNA synthesis was performed using the FastKing RT kit (Tiangen Biotech Co., Ltd, KR118). qPCR was performed using SYBR Green PCR Material Mixture reagent (Tiangen Biotech Co., Ltd, FP205) on the Real-time PCR Detection System (Bio-Rad), with primer sequences shown in Table S1. .. The following antibodies were used: RNF8 (Proteintech, #14112-1-AP), β-actin (Affinity, #AF7018), Tri-Methyl-Histone H3 (Lys27) (Affinity, #DF6941), IL6 (Affinity, #DF6087), IFN-γ (Affinity, #DF6045), γ-H2AX (Servicebio, # GB111841-100), CD86 (Affinity, #DF6332), CD74 (Immunoway, #YT5464), CD163 (Proteintech, #16646-1-AP), OPTN (Proteintech, #10837-1-AP), CD206 (Affinity, #DF4149), CD80 (Proteintech, #66406-1-Ig), p62 (Proteintech, #31403-1-AP), LC3(Proteintech, #14600-1-AP), c-kit(Affinity, #AF6153), Scp3(Proteintech, #23024-1-AP), Flag (Proteintech, #20543- 1-AP), HA (Proteintech, #66006-2-Ig), His (Proteintech, #66005-1-Ig), ubiquitin (Cell Signaling Technology, #91112S), K48 (Cell Signaling Technology, #12805), K63 (Cell Signaling Technology, #5621S). ..

    Article Title: The RNF8/OPTN/KDM6A axis controls macrophage polarization to maintain testicular microenvironment homeostasis
    Article Snippet: Total RNA was extracted from testicular tissue and 293T cells using TRIZOL (Ambient, 95141801), and RNA concentration and purity were detected using nanodrop spectrophotometry (Thermo Scientific). cDNA synthesis was performed using the FastKing RT kit (Tiangen Biotech Co., Ltd, KR118). qPCR was performed using SYBR Green PCR Material Mixture reagent (Tiangen Biotech Co., Ltd, FP205) on the Real-time PCR Detection System (Bio-Rad), with primer sequences shown in Table S1. .. The following antibodies were used: RNF8 (Proteintech, #14112-1-AP), β-actin (Affinity, #AF7018), Tri-Methyl-Histone H3 (Lys27) (Affinity, #DF6941), IL6 (Affinity, #DF6087), IFN-γ (Affinity, #DF6045), γ-H2AX (Servicebio, # GB111841-100), CD86 (Affinity, #DF6332), CD74 (Immunoway, #YT5464), CD163 (Proteintech, #16646-1-AP), OPTN (Proteintech, #10837-1-AP), CD206 (Affinity, #DF4149), CD80 (Proteintech, #66406-1-Ig), p62 (Proteintech, #31403-1-AP), LC3(Proteintech, #14600-1-AP), c-kit(Affinity, #AF6153), Scp3(Proteintech, #23024-1-AP), Flag (Proteintech, #20543-1-AP), HA (Proteintech, #66006-2-Ig), His (Proteintech, #66005-1-Ig), ubiquitin (Cell Signaling Technology, #91112S), K48 (Cell Signaling Technology, #12805), K63 (Cell Signaling Technology, #5621S). ..

    Western Blot:

    Article Title: AMPK–mTOR–Mediated Activation of Autophagy Promotes Formation of Dormant Polyploid Giant Cancer Cells
    Article Snippet: .. Antibodies used for IF, western blot, and IHC Antibodies Manufacturer Catalog number Dilution WB IHC IF Cyclin D1 Abcam Ab134175 1:1000 Cyclin B1 Proteintech 55004-1-AP 1:600 Cyclin E1 Proteintech 11554-1-AP 1:800 CDK8 Proteintech 22067-1-AP 1:600 CD133 Proteintech 66666-1-lg 1:500 P-gp Proteintech 22336-1-AP 1:600 ACOX-1 Proteintech 10957-1-AP 1:800 FASN Proteintech 10624-2-AP 1:800 HIF-1a Proteintech 20960-1-AP 1:600 EIF4A-1 Abcam Ab31217 1:1000 ATP-5A Proteintech 66037-1-lg 1:800 1:100 COX4IA Sangon Biotech D262690-0025 1:1000 1:100 ATG5 Cell Signaling Technology (CST) 12994T 1:1000 ATG7 Abcam Ab52472 1:1000 ATG16L1 Proteintech 19812-1-AP 1:500 ATG12 Proteintech 11264-1-AP 1:500 LC3 Proteintech 14600-1-AP 1:1000 1:500 1:200 P62 Proteintech 18420-1-AP 1:800 1:100 p-AKT CST 4060T 1:1000 t-AKT CST 4691T 1:1000 p-MEK CST 9154S 1:1000 t-MEK CST 4694S 1:1000 p-AMPK CST 2531S 1:1000 t-AMPK CST 2532S 1:1000 27 p-MTOR CST 5536T 1:1000 t-MTOR CST 2983T 1:1000 TOM20 Santa SC-17764 1:50 Tublin Santa SC-5286 1:50 SIRT1 Proteintech 13161-1-AP 1:600 LKB1 Proteintech 10746-1-AP 1:600 RIPK1 ABclonal A19580 1:1000 pS166 RIPK1 Proteintech 28252-1-AP 1:600 pS320 RIPK1 Dia-An Biotechnology Co., Ltd custom antibody 1:300 p-p38 ABclonal AP0526 1:1000 p38 ABclonal A14401 1:1000 H3K27ac Abcam ab4729 GAPDH Proteintech 10494-1-AP 1:2000 28 Supplementary Table S3. ..

    Article Title: IGF2BP1-HAX-1 positive feedback loop-mediated HAX-1 overexpression blocks autophagic flux and promotes chemoresistance in nasopharyngeal carcinoma
    Article Snippet: .. Antibodies Manufacturer Catalog number Dilution WB IHC IF HAX-1 ABclonal A5551 1:500 1:100 1:100 ABCC1 ABclonal A2223 1:500 1:100 P glycoprotein proteintech 22336-1-AP 1:500 1:100 KI67 proteintech 27309-1-AP 1:2,000 SQSTM1 proteintech 18420-1-AP 1:1,000 1:500 Beclin 1 proteintech 11306-1-AP 1:1,000 ATG5 proteintech 10181-2-AP 1:500 LC3 proteintech 14600-1-AP 1:500 NBR1 proteintech 16004-1-AP 1:500 OPTN proteintech 10837-1-AP 1:500 TAX1BP1 proteintech 14424-1-AP 1:500 Rab7a proteintech 55469-1-AP 1:500 1:100 1:20 Lamp1 Santa Cruz Biotechnology sc-20011 1:10 SNAP29 proteintech 12704-1-AP 1:200 1:10 VAMP8 proteintech 15546-1-AP 1:200 1:10 STX17 proteintech 17815-1-AP 1:200 IGF2BP1 proteintech 22803-1-AP 1:500 1:100 22 NEDD4 proteintech 21698-1-AP 1:500 1:50 HA-Tag Cell Signaling Technology #3724 1:500 Myc-Tag Cell Signaling Technology #2276 1:500 YTHDF1 proteintech 17479-1-AP 1:1,000 YTHDF2 proteintech 24744-1-AP 1:1,000 WTAP proteintech 60188-1-Ig 1:2,000 GAPDH proteintech 10494-1-AP 1:2,000 23 Table S3. ..

    Immunohistochemistry:

    Article Title: AMPK–mTOR–Mediated Activation of Autophagy Promotes Formation of Dormant Polyploid Giant Cancer Cells
    Article Snippet: .. Antibodies used for IF, western blot, and IHC Antibodies Manufacturer Catalog number Dilution WB IHC IF Cyclin D1 Abcam Ab134175 1:1000 Cyclin B1 Proteintech 55004-1-AP 1:600 Cyclin E1 Proteintech 11554-1-AP 1:800 CDK8 Proteintech 22067-1-AP 1:600 CD133 Proteintech 66666-1-lg 1:500 P-gp Proteintech 22336-1-AP 1:600 ACOX-1 Proteintech 10957-1-AP 1:800 FASN Proteintech 10624-2-AP 1:800 HIF-1a Proteintech 20960-1-AP 1:600 EIF4A-1 Abcam Ab31217 1:1000 ATP-5A Proteintech 66037-1-lg 1:800 1:100 COX4IA Sangon Biotech D262690-0025 1:1000 1:100 ATG5 Cell Signaling Technology (CST) 12994T 1:1000 ATG7 Abcam Ab52472 1:1000 ATG16L1 Proteintech 19812-1-AP 1:500 ATG12 Proteintech 11264-1-AP 1:500 LC3 Proteintech 14600-1-AP 1:1000 1:500 1:200 P62 Proteintech 18420-1-AP 1:800 1:100 p-AKT CST 4060T 1:1000 t-AKT CST 4691T 1:1000 p-MEK CST 9154S 1:1000 t-MEK CST 4694S 1:1000 p-AMPK CST 2531S 1:1000 t-AMPK CST 2532S 1:1000 27 p-MTOR CST 5536T 1:1000 t-MTOR CST 2983T 1:1000 TOM20 Santa SC-17764 1:50 Tublin Santa SC-5286 1:50 SIRT1 Proteintech 13161-1-AP 1:600 LKB1 Proteintech 10746-1-AP 1:600 RIPK1 ABclonal A19580 1:1000 pS166 RIPK1 Proteintech 28252-1-AP 1:600 pS320 RIPK1 Dia-An Biotechnology Co., Ltd custom antibody 1:300 p-p38 ABclonal AP0526 1:1000 p38 ABclonal A14401 1:1000 H3K27ac Abcam ab4729 GAPDH Proteintech 10494-1-AP 1:2000 28 Supplementary Table S3. ..

    Article Title: IGF2BP1-HAX-1 positive feedback loop-mediated HAX-1 overexpression blocks autophagic flux and promotes chemoresistance in nasopharyngeal carcinoma
    Article Snippet: .. Antibodies Manufacturer Catalog number Dilution WB IHC IF HAX-1 ABclonal A5551 1:500 1:100 1:100 ABCC1 ABclonal A2223 1:500 1:100 P glycoprotein proteintech 22336-1-AP 1:500 1:100 KI67 proteintech 27309-1-AP 1:2,000 SQSTM1 proteintech 18420-1-AP 1:1,000 1:500 Beclin 1 proteintech 11306-1-AP 1:1,000 ATG5 proteintech 10181-2-AP 1:500 LC3 proteintech 14600-1-AP 1:500 NBR1 proteintech 16004-1-AP 1:500 OPTN proteintech 10837-1-AP 1:500 TAX1BP1 proteintech 14424-1-AP 1:500 Rab7a proteintech 55469-1-AP 1:500 1:100 1:20 Lamp1 Santa Cruz Biotechnology sc-20011 1:10 SNAP29 proteintech 12704-1-AP 1:200 1:10 VAMP8 proteintech 15546-1-AP 1:200 1:10 STX17 proteintech 17815-1-AP 1:200 IGF2BP1 proteintech 22803-1-AP 1:500 1:100 22 NEDD4 proteintech 21698-1-AP 1:500 1:50 HA-Tag Cell Signaling Technology #3724 1:500 Myc-Tag Cell Signaling Technology #2276 1:500 YTHDF1 proteintech 17479-1-AP 1:1,000 YTHDF2 proteintech 24744-1-AP 1:1,000 WTAP proteintech 60188-1-Ig 1:2,000 GAPDH proteintech 10494-1-AP 1:2,000 23 Table S3. ..

    other:

    Article Title: RPL11 promotes non-small cell lung cancer cell proliferation via regulating endoplasmic reticulum stress and cell autophagy
    Article Snippet: Primary and secondary antibodies used are list as follows: RPL11 (Proteintech, 16277-1- AP), Cyclin D1 (Proteintech, 26939-1-AP), BIP/GRP78 (Proteintech, 66574-1-Ig), ATF4 (Proteintech, 10835- 1-AP), CHOP (Proteintech, 15204-1-AP), LC3 (Proteintech, 14600-1-AP), Beclin1 (BOSTER, BM5181), p62 (BOSTER, M00300-1), CDK4 (Proteintech, 11026-1-AP), FLAG (Proteintech, 66008-3-1g), phospho-IRE1 (A nity, AF7150), IRE1 (A nity, DF7709), β-actin (Proteintech, 66009-1-Ig).

    Chromatin Immunoprecipitation:

    Article Title: Autophagy enhanced by curcumin ameliorates inflammation in atherogenesis via the TFEB–P300–BRD4 axis
    Article Snippet: .. Factor or PTM Vendor Cat. No. Dilution for blotting Vol. for ChIP (μL) P62 Cell Signaling Technology 5114S 1:1,000 LC3 Proteintech 14600-1-AP 1:1,000 Actin Biogot AP0060 1:1,000 TFEB Santa sc-166736 1:1,000 10 TFEB Proteintech 13372-1-AP 1:1,000 p-TFEB Affinity AF3708 1:1,000 PCNA Cell Signaling Technology 13110 1:1,000 ATG5 Cell Signaling Technology 12994 1:1,000 P300 Abcam ab54984 1:1,000 P300 Bethyl A300-358A 10 BRD2 Santa sc-393720 1:1,000 BRD2 Cell Signaling Technology 5848S 10 BRD3 Proteintech 11859-1-AP 1:1,000 8 BRD4 Abcam ab243862 1:1,000 10 Histone 3 Abcam ab1791 1:1,000 8 mTOR Cell Signaling Technology 2972S 1:1,000 p-mTOR Cell Signaling Technology 2971S 1:1,000 Acetylated histone 3 Abcam ab4729 1:1,000 MED1 Bethyl A300-793A 1:1,000 5 AKT Cell Signaling Technology 9272 1:1,000 p-AKT Cell Signaling Technology 4060S 1:1,000 Calcineurin Cell Signaling Technology 2614 1:1,000 S6K1 Cell Signaling Technology 34475 1:1,000 p-S6K1 Cell Signaling Technology 97596 1:1,000 H3K27ac Abcam ab177178 1:1,000 8 H3K4me1 Abcam ab176877 1:1,000 H3K27me3 Abcam ab6002 1:1,000 H3K9ac Abcam ab32129 1:1,000 H3K14ac Abcam ab52946 1:1,000 H3K18ac Abcam ab177870 1:1,000 H3K23ac Abcam ab177275 1:1,000 BTK Proteintech 21581-1-AP 1:1,000 p-BTK Cell Signaling Technology 5082 1:1,000 HRP-conjugated goat anti-rabbit secondary antibody Jackson Labs 111035003 1:30,000 HRP-conjugated goat antimouse secondary antibody Jackson Labs 115036003 1:30,000 Table S3 RT-PCR primers used in this study. ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Autophagy enhanced by curcumin ameliorates inflammation in atherogenesis via the TFEB–P300–BRD4 axis
    Article Snippet: .. Factor or PTM Vendor Cat. No. Dilution for blotting Vol. for ChIP (μL) P62 Cell Signaling Technology 5114S 1:1,000 LC3 Proteintech 14600-1-AP 1:1,000 Actin Biogot AP0060 1:1,000 TFEB Santa sc-166736 1:1,000 10 TFEB Proteintech 13372-1-AP 1:1,000 p-TFEB Affinity AF3708 1:1,000 PCNA Cell Signaling Technology 13110 1:1,000 ATG5 Cell Signaling Technology 12994 1:1,000 P300 Abcam ab54984 1:1,000 P300 Bethyl A300-358A 10 BRD2 Santa sc-393720 1:1,000 BRD2 Cell Signaling Technology 5848S 10 BRD3 Proteintech 11859-1-AP 1:1,000 8 BRD4 Abcam ab243862 1:1,000 10 Histone 3 Abcam ab1791 1:1,000 8 mTOR Cell Signaling Technology 2972S 1:1,000 p-mTOR Cell Signaling Technology 2971S 1:1,000 Acetylated histone 3 Abcam ab4729 1:1,000 MED1 Bethyl A300-793A 1:1,000 5 AKT Cell Signaling Technology 9272 1:1,000 p-AKT Cell Signaling Technology 4060S 1:1,000 Calcineurin Cell Signaling Technology 2614 1:1,000 S6K1 Cell Signaling Technology 34475 1:1,000 p-S6K1 Cell Signaling Technology 97596 1:1,000 H3K27ac Abcam ab177178 1:1,000 8 H3K4me1 Abcam ab176877 1:1,000 H3K27me3 Abcam ab6002 1:1,000 H3K9ac Abcam ab32129 1:1,000 H3K14ac Abcam ab52946 1:1,000 H3K18ac Abcam ab177870 1:1,000 H3K23ac Abcam ab177275 1:1,000 BTK Proteintech 21581-1-AP 1:1,000 p-BTK Cell Signaling Technology 5082 1:1,000 HRP-conjugated goat anti-rabbit secondary antibody Jackson Labs 111035003 1:30,000 HRP-conjugated goat antimouse secondary antibody Jackson Labs 115036003 1:30,000 Table S3 RT-PCR primers used in this study. ..



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    Proteintech lc3 14600 1 ap proteintech wuhan china
    Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for <t>LC3B</t> (green). Quantification of the mean fluorescence intensity of <t>LC3</t> is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001
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    Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for <t>LC3B</t> (green). Quantification of the mean fluorescence intensity of <t>LC3</t> is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001
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    Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for <t>LC3B</t> (green). Quantification of the mean fluorescence intensity of <t>LC3</t> is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001
    1 Ap Proteintech, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for <t>LC3B</t> (green). Quantification of the mean fluorescence intensity of <t>LC3</t> is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001
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    Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for <t>LC3B</t> (green). Quantification of the mean fluorescence intensity of <t>LC3</t> is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001
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    Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for <t>LC3B</t> (green). Quantification of the mean fluorescence intensity of <t>LC3</t> is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001
    1 Ap Proteintech Ab 2137737, supplied by Proteintech, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for LC3B (green). Quantification of the mean fluorescence intensity of LC3 is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001

    Journal: The Journal of Headache and Pain

    Article Title: S100A11 regulates microglial inflammatory response in neuropathic pain via H3K27ac-TFEB-mitochondrial autophagy axis

    doi: 10.1186/s10194-026-02328-9

    Figure Lengend Snippet: Knockdown of S100A11 improves inflammatory response and pyroptosis in primary microglia by inhibiting mitophagy. A Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; B-E Relative levels normalized to COXIV were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). G-H Labeling of mitochondria in primary microglia with Tom20 (red); immunofluorescence staining for LC3B (green). Quantification of the mean fluorescence intensity of LC3 is shown in panels F and I . (Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Immunofluorescence staining of CD68 ( J ) and CD206 ( K ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels. L , M ( Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with appropriate post hoc tests), for three independent experiments. Q Western blot analysis of NLRP3 protein levels of microglia; N Relative levels normalized to β-actin was quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Immunofluorescence staining of NLRP3 ( R ) and Caspase1 ( S ) in microglia (red); green fluorescence indicates phalloidin. Quantification of relative fluorescence intensities is shown in panels O , P (Statistical significance was determined by one-way ANOVA with appropriate post hoc tests) for three independent experiments. Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001

    Article Snippet: Membranes were blocked with 5% skim milk for 60 min, rinsed with TBST, and incubated with primary antibodies at 4 °C overnight (β-actin: Proteintech, 66009-1-Ig, 1:10000; S100A11: Abcam, ab169530, 1:1000; NLRP3: AdipoGen, AG-20B-0014, 1:1000; GSDMD-N: Abcam, ab215203, 1:1000; ASC: Wanleibio, WL02462, 1:500; Caspase1: CST, 89332, 1:1000; Bax: Proteintech, 50599-2-Ig, 1:2000; Bcl-2: abclone, A19693, 1:2000; COXIV: Proteintech, 11242-1-AP, 1:1000; LC3: Proteintech, 14600-1-AP, 1:2000; P62: Proteintech, 18420-1-AP, 1:2000; Parkin: Proteintech, 14060-1-AP, 1:1000; Pink1: Proteintech, 23274-1-AP, 1:2000; H3: Proteintech, 17168-1-AP, 1:2000; TFEB: Proteintech, 13372-1-AP, 1:1000; TFEC: Proteintech, 13547-1-AP, 1:1000; TFE3: Proteintech, 14480-1-AP, 1:1000; H3K9ac: CST, 9649, 1:1000; H3K18ac: CST, 13998, 1:1000; H3K27ac: CST, 8173, 1:1000; H4K12ac; CST, 13944, 1:1000; H4K16ac: CST, 13534, 1:1000).

    Techniques: Knockdown, Western Blot, Labeling, Immunofluorescence, Staining, Fluorescence

    S100A11 knockdown alleviates microglial inflammation by inhibiting mitophagy through the TFEB/H3K27ac axis. A-B Western blot analysis of TFEB, TFEC, and TFE3 protein expression levels in the cytoplasm and nucleus of primary microglia, along with the corresponding detection and quantification plots ( D-F , for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). C Immunofluorescence staining of TFEB in primary microglia. G Western blot analysis of pink1, Parkin, P62, LC3II protein levels of primary microglia; H-K Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). M Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green). Quantification of the mean fluorescence intensity of LC3 is shown in panel L . and relative fluorescence intensity was quantified in panel L ( n = 3). N Western blot analysis of H4K16ac, H4K12ac, H3K27ac, H3K18ac, H3K9ac protein levels of microglia; O-S Relative levels normalized to histone H3 were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). T Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; U-X Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Z Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green), and relative fluorescence intensity was quantified in panel Y ( n = 3). Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001

    Journal: The Journal of Headache and Pain

    Article Title: S100A11 regulates microglial inflammatory response in neuropathic pain via H3K27ac-TFEB-mitochondrial autophagy axis

    doi: 10.1186/s10194-026-02328-9

    Figure Lengend Snippet: S100A11 knockdown alleviates microglial inflammation by inhibiting mitophagy through the TFEB/H3K27ac axis. A-B Western blot analysis of TFEB, TFEC, and TFE3 protein expression levels in the cytoplasm and nucleus of primary microglia, along with the corresponding detection and quantification plots ( D-F , for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). C Immunofluorescence staining of TFEB in primary microglia. G Western blot analysis of pink1, Parkin, P62, LC3II protein levels of primary microglia; H-K Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). M Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green). Quantification of the mean fluorescence intensity of LC3 is shown in panel L . and relative fluorescence intensity was quantified in panel L ( n = 3). N Western blot analysis of H4K16ac, H4K12ac, H3K27ac, H3K18ac, H3K9ac protein levels of microglia; O-S Relative levels normalized to histone H3 were quantified (for three independent experiments, statistical significance was determined by two-way ANOVA with appropriate post hoc tests). T Western blot analysis of Pink1, Parkin, P62, LC3II protein levels of primary microglia; U-X Relative levels normalized to COX IV were quantified (for three independent experiments, statistical significance was determined by one-way ANOVA with appropriate post hoc tests). Z Labeling of mitochondria (red) in primary microglia with Tom20; immunofluorescence staining for LC3B (green), and relative fluorescence intensity was quantified in panel Y ( n = 3). Scale bars as indicated. Data are presented as mean ± SEM. * P < 0.05 , ** P < 0.01 , *** P < 0.001

    Article Snippet: Membranes were blocked with 5% skim milk for 60 min, rinsed with TBST, and incubated with primary antibodies at 4 °C overnight (β-actin: Proteintech, 66009-1-Ig, 1:10000; S100A11: Abcam, ab169530, 1:1000; NLRP3: AdipoGen, AG-20B-0014, 1:1000; GSDMD-N: Abcam, ab215203, 1:1000; ASC: Wanleibio, WL02462, 1:500; Caspase1: CST, 89332, 1:1000; Bax: Proteintech, 50599-2-Ig, 1:2000; Bcl-2: abclone, A19693, 1:2000; COXIV: Proteintech, 11242-1-AP, 1:1000; LC3: Proteintech, 14600-1-AP, 1:2000; P62: Proteintech, 18420-1-AP, 1:2000; Parkin: Proteintech, 14060-1-AP, 1:1000; Pink1: Proteintech, 23274-1-AP, 1:2000; H3: Proteintech, 17168-1-AP, 1:2000; TFEB: Proteintech, 13372-1-AP, 1:1000; TFEC: Proteintech, 13547-1-AP, 1:1000; TFE3: Proteintech, 14480-1-AP, 1:1000; H3K9ac: CST, 9649, 1:1000; H3K18ac: CST, 13998, 1:1000; H3K27ac: CST, 8173, 1:1000; H4K12ac; CST, 13944, 1:1000; H4K16ac: CST, 13534, 1:1000).

    Techniques: Knockdown, Western Blot, Expressing, Immunofluorescence, Staining, Labeling, Fluorescence