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coil myosin like bcl2  (Proteintech)


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    Proteintech coil myosin like bcl2
    rtPA attenuates neurological behavior impairment and apoptosis after ICH. (A–C) Left forelimb placement experiment, corner turn experiment, and modified Garcia score testing were conducted at 1 hour before surgery and 6, 24, and 72 hours after surgery ( n = 14 per group). (D) H&E staining (top) and Nissl staining (bottom) of peri-hematoma tissue at 72 hours after ICH and rtPA treatments ( n = 3 per group). Scale bars: 100 µm. (E) Representative picture of TUNEL staining of peri-hematoma tissue conducted at 72 hours after ICH and rtPA treatments ( n = 3–6 per group). Scale bars: 100 µm. (F) The proportion of TUNEL-positive cells to all nucleated cells surrounding the hematoma ( n = 3–6 per group). (G–J) Analysis of apoptosis-associated proteins at 24 and 72 hours after treatment ( n = 3). Data are represented as mean ± SEM. * P < 0.05, **** P < 0.0001, vs. sham group; &P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs . ICH + vehicle group (two-way analysis of variance followed by Bonferroni post hoc test (A–C) or one-way analysis of variance followed by Tukey’s post hoc test (F, I, J). bax: Apoptosis regulator bax; <t>bcl2:</t> apoptosis regulator bcl2; DAPI: 4′,6-diamidino-2-phenylindole; ER: endoplasmic reticulum; H&E staining: hematoxylin & eosin staining; ICH: intracerebral hemorrhage; rtPA: recombinant tissue plasminogen activator; TUNEL: terminal deoxynucleotidyl transferase dUTP nick-end labeling.
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    Images

    1) Product Images from "Recombinant tissue plasminogen activator protects neurons after intracerebral hemorrhage through activating the PI3K/AKT/mTOR pathway"

    Article Title: Recombinant tissue plasminogen activator protects neurons after intracerebral hemorrhage through activating the PI3K/AKT/mTOR pathway

    Journal: Neural Regeneration Research

    doi: 10.4103/NRR.NRR-D-23-01953

    rtPA attenuates neurological behavior impairment and apoptosis after ICH. (A–C) Left forelimb placement experiment, corner turn experiment, and modified Garcia score testing were conducted at 1 hour before surgery and 6, 24, and 72 hours after surgery ( n = 14 per group). (D) H&E staining (top) and Nissl staining (bottom) of peri-hematoma tissue at 72 hours after ICH and rtPA treatments ( n = 3 per group). Scale bars: 100 µm. (E) Representative picture of TUNEL staining of peri-hematoma tissue conducted at 72 hours after ICH and rtPA treatments ( n = 3–6 per group). Scale bars: 100 µm. (F) The proportion of TUNEL-positive cells to all nucleated cells surrounding the hematoma ( n = 3–6 per group). (G–J) Analysis of apoptosis-associated proteins at 24 and 72 hours after treatment ( n = 3). Data are represented as mean ± SEM. * P < 0.05, **** P < 0.0001, vs. sham group; &P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs . ICH + vehicle group (two-way analysis of variance followed by Bonferroni post hoc test (A–C) or one-way analysis of variance followed by Tukey’s post hoc test (F, I, J). bax: Apoptosis regulator bax; bcl2: apoptosis regulator bcl2; DAPI: 4′,6-diamidino-2-phenylindole; ER: endoplasmic reticulum; H&E staining: hematoxylin & eosin staining; ICH: intracerebral hemorrhage; rtPA: recombinant tissue plasminogen activator; TUNEL: terminal deoxynucleotidyl transferase dUTP nick-end labeling.
    Figure Legend Snippet: rtPA attenuates neurological behavior impairment and apoptosis after ICH. (A–C) Left forelimb placement experiment, corner turn experiment, and modified Garcia score testing were conducted at 1 hour before surgery and 6, 24, and 72 hours after surgery ( n = 14 per group). (D) H&E staining (top) and Nissl staining (bottom) of peri-hematoma tissue at 72 hours after ICH and rtPA treatments ( n = 3 per group). Scale bars: 100 µm. (E) Representative picture of TUNEL staining of peri-hematoma tissue conducted at 72 hours after ICH and rtPA treatments ( n = 3–6 per group). Scale bars: 100 µm. (F) The proportion of TUNEL-positive cells to all nucleated cells surrounding the hematoma ( n = 3–6 per group). (G–J) Analysis of apoptosis-associated proteins at 24 and 72 hours after treatment ( n = 3). Data are represented as mean ± SEM. * P < 0.05, **** P < 0.0001, vs. sham group; &P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs . ICH + vehicle group (two-way analysis of variance followed by Bonferroni post hoc test (A–C) or one-way analysis of variance followed by Tukey’s post hoc test (F, I, J). bax: Apoptosis regulator bax; bcl2: apoptosis regulator bcl2; DAPI: 4′,6-diamidino-2-phenylindole; ER: endoplasmic reticulum; H&E staining: hematoxylin & eosin staining; ICH: intracerebral hemorrhage; rtPA: recombinant tissue plasminogen activator; TUNEL: terminal deoxynucleotidyl transferase dUTP nick-end labeling.

    Techniques Used: Modification, Staining, TUNEL Assay, Recombinant

    rtPA attenuates autophagy in peri-hematoma tissue after ICH. (A–D) Analysis of autophagy-associated proteins of peri-hematoma tissue at 24 hours after treatment. (E–H) Analysis of autophagy-associated proteins of peri-hematoma tissue at 72 hours after treatment. Data are shown as mean ± SEM ( n = 3 per group). ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . sham group; & P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, #### P < 0.0001, vs . ICH + vehicle group (one-way analysis of variance followed by Tukey’s post hoc test). beclin1: Coiled-coil myosin-like bcl2-interacting protein; ICH: intracerebral hemorrhage; LC3: microtubule-associated proteins 1A/1B light chain 3B; p62: sequestosome-1/ubiquitin-binding protein p62; rtPA: recombinant tissue plasminogen activator.
    Figure Legend Snippet: rtPA attenuates autophagy in peri-hematoma tissue after ICH. (A–D) Analysis of autophagy-associated proteins of peri-hematoma tissue at 24 hours after treatment. (E–H) Analysis of autophagy-associated proteins of peri-hematoma tissue at 72 hours after treatment. Data are shown as mean ± SEM ( n = 3 per group). ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . sham group; & P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, #### P < 0.0001, vs . ICH + vehicle group (one-way analysis of variance followed by Tukey’s post hoc test). beclin1: Coiled-coil myosin-like bcl2-interacting protein; ICH: intracerebral hemorrhage; LC3: microtubule-associated proteins 1A/1B light chain 3B; p62: sequestosome-1/ubiquitin-binding protein p62; rtPA: recombinant tissue plasminogen activator.

    Techniques Used: Ubiquitin Proteomics, Binding Assay, Recombinant

    rtPA attenuates neuron apoptosis and autophagy after experimental ICH in vitro . (A–C) The DEGs between control group and hemin group associated with autophagy animals (KEGG: mmu04140), positive regulation of neuron apoptotic process (GO: 0043525), and positive regulation of response to endoplasmic reticulum stress (GO: 1905898) were screened, and the transcriptional levels of DEGs in each group are presented as heatmaps. (D, E) Analysis of apoptosis-associated proteins. (F) Transmission electron microscopy images of neurons after hemin and rtPA treatment. Red asterisk indicates the autophagosome, black arrows indicate the endoplasmic reticulum, and N means nucleus. Scale bars: 1 µm. (G–J) Analysis of autophagy-associated proteins. Data are shown as mean ± SEM ( n = 3–4). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs. hemin group (one-way analysis of variance followed by Tukey’s post hoc test). bax: Apoptosis regulator bax; bcl2: apoptosis regulator bcl2; beclin1: coiled-coil myosin-like bcl2-interacting protein; DEGs: differential expression genes; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; LC3: microtubule-associated proteins 1A/1B light chain 3B; p62: sequestosome-1/ubiquitin-binding protein p62; rtPA: recombinant tissue plasminogen activator.
    Figure Legend Snippet: rtPA attenuates neuron apoptosis and autophagy after experimental ICH in vitro . (A–C) The DEGs between control group and hemin group associated with autophagy animals (KEGG: mmu04140), positive regulation of neuron apoptotic process (GO: 0043525), and positive regulation of response to endoplasmic reticulum stress (GO: 1905898) were screened, and the transcriptional levels of DEGs in each group are presented as heatmaps. (D, E) Analysis of apoptosis-associated proteins. (F) Transmission electron microscopy images of neurons after hemin and rtPA treatment. Red asterisk indicates the autophagosome, black arrows indicate the endoplasmic reticulum, and N means nucleus. Scale bars: 1 µm. (G–J) Analysis of autophagy-associated proteins. Data are shown as mean ± SEM ( n = 3–4). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs. hemin group (one-way analysis of variance followed by Tukey’s post hoc test). bax: Apoptosis regulator bax; bcl2: apoptosis regulator bcl2; beclin1: coiled-coil myosin-like bcl2-interacting protein; DEGs: differential expression genes; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; LC3: microtubule-associated proteins 1A/1B light chain 3B; p62: sequestosome-1/ubiquitin-binding protein p62; rtPA: recombinant tissue plasminogen activator.

    Techniques Used: In Vitro, Control, Transmission Assay, Electron Microscopy, Quantitative Proteomics, Ubiquitin Proteomics, Binding Assay, Recombinant



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    rtPA attenuates neurological behavior impairment and apoptosis after ICH. (A–C) Left forelimb placement experiment, corner turn experiment, and modified Garcia score testing were conducted at 1 hour before surgery and 6, 24, and 72 hours after surgery ( n = 14 per group). (D) H&E staining (top) and Nissl staining (bottom) of peri-hematoma tissue at 72 hours after ICH and rtPA treatments ( n = 3 per group). Scale bars: 100 µm. (E) Representative picture of TUNEL staining of peri-hematoma tissue conducted at 72 hours after ICH and rtPA treatments ( n = 3–6 per group). Scale bars: 100 µm. (F) The proportion of TUNEL-positive cells to all nucleated cells surrounding the hematoma ( n = 3–6 per group). (G–J) Analysis of apoptosis-associated proteins at 24 and 72 hours after treatment ( n = 3). Data are represented as mean ± SEM. * P < 0.05, **** P < 0.0001, vs. sham group; &P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs . ICH + vehicle group (two-way analysis of variance followed by Bonferroni post hoc test (A–C) or one-way analysis of variance followed by Tukey’s post hoc test (F, I, J). bax: Apoptosis regulator bax; <t>bcl2:</t> apoptosis regulator bcl2; DAPI: 4′,6-diamidino-2-phenylindole; ER: endoplasmic reticulum; H&E staining: hematoxylin & eosin staining; ICH: intracerebral hemorrhage; rtPA: recombinant tissue plasminogen activator; TUNEL: terminal deoxynucleotidyl transferase dUTP nick-end labeling.
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    Image Search Results


    rtPA attenuates neurological behavior impairment and apoptosis after ICH. (A–C) Left forelimb placement experiment, corner turn experiment, and modified Garcia score testing were conducted at 1 hour before surgery and 6, 24, and 72 hours after surgery ( n = 14 per group). (D) H&E staining (top) and Nissl staining (bottom) of peri-hematoma tissue at 72 hours after ICH and rtPA treatments ( n = 3 per group). Scale bars: 100 µm. (E) Representative picture of TUNEL staining of peri-hematoma tissue conducted at 72 hours after ICH and rtPA treatments ( n = 3–6 per group). Scale bars: 100 µm. (F) The proportion of TUNEL-positive cells to all nucleated cells surrounding the hematoma ( n = 3–6 per group). (G–J) Analysis of apoptosis-associated proteins at 24 and 72 hours after treatment ( n = 3). Data are represented as mean ± SEM. * P < 0.05, **** P < 0.0001, vs. sham group; &P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs . ICH + vehicle group (two-way analysis of variance followed by Bonferroni post hoc test (A–C) or one-way analysis of variance followed by Tukey’s post hoc test (F, I, J). bax: Apoptosis regulator bax; bcl2: apoptosis regulator bcl2; DAPI: 4′,6-diamidino-2-phenylindole; ER: endoplasmic reticulum; H&E staining: hematoxylin & eosin staining; ICH: intracerebral hemorrhage; rtPA: recombinant tissue plasminogen activator; TUNEL: terminal deoxynucleotidyl transferase dUTP nick-end labeling.

    Journal: Neural Regeneration Research

    Article Title: Recombinant tissue plasminogen activator protects neurons after intracerebral hemorrhage through activating the PI3K/AKT/mTOR pathway

    doi: 10.4103/NRR.NRR-D-23-01953

    Figure Lengend Snippet: rtPA attenuates neurological behavior impairment and apoptosis after ICH. (A–C) Left forelimb placement experiment, corner turn experiment, and modified Garcia score testing were conducted at 1 hour before surgery and 6, 24, and 72 hours after surgery ( n = 14 per group). (D) H&E staining (top) and Nissl staining (bottom) of peri-hematoma tissue at 72 hours after ICH and rtPA treatments ( n = 3 per group). Scale bars: 100 µm. (E) Representative picture of TUNEL staining of peri-hematoma tissue conducted at 72 hours after ICH and rtPA treatments ( n = 3–6 per group). Scale bars: 100 µm. (F) The proportion of TUNEL-positive cells to all nucleated cells surrounding the hematoma ( n = 3–6 per group). (G–J) Analysis of apoptosis-associated proteins at 24 and 72 hours after treatment ( n = 3). Data are represented as mean ± SEM. * P < 0.05, **** P < 0.0001, vs. sham group; &P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs . ICH + vehicle group (two-way analysis of variance followed by Bonferroni post hoc test (A–C) or one-way analysis of variance followed by Tukey’s post hoc test (F, I, J). bax: Apoptosis regulator bax; bcl2: apoptosis regulator bcl2; DAPI: 4′,6-diamidino-2-phenylindole; ER: endoplasmic reticulum; H&E staining: hematoxylin & eosin staining; ICH: intracerebral hemorrhage; rtPA: recombinant tissue plasminogen activator; TUNEL: terminal deoxynucleotidyl transferase dUTP nick-end labeling.

    Article Snippet: The following primary antibodies were used for analysis: bcl2 (rabbit, 1:1000, Proteintech, Cat# 12789-1-AP, RRID: AB_2227948), bax (rabbit, 1:1000, Proteintech, Cat# 50599-2-Ig, RRID: AB_2061561), coiled-coil myosin-like bcl2-interacting protein (beclin1; rabbit, 1:1000, Proteintech, Cat# 11306-1-AP, RRID: AB_2259061), sequestosome-1/ubiquitin-binding protein p62 (SQSTM1/p62; rabbit, 1:1000, Abclonal, Cat# A11250, RRID: AB_2758477), microtubule-associated proteins 1A/1B light chain 3B (LC3; rabbit, 1:1000, Abcam, Cat# ab48394, RRID: AB_881433), endoplasmic reticulum chaperone BiP (Grp78/BIP; mouse, 1:1000, Proteintech, Cat# 66574-1-Ig, RRID: AB_2881934), cyclic AMP-dependent transcription factor ATF-6 alpha (ATF6; rabbit, 1:1000, Proteintech, Cat# 24169-1-AP, RRID: AB_2876891), PRKR-like endoplasmic reticulum kinase (PERK; rabbit, 1:1000, Cell Signaling Technology, Cat# 3192S, RRID: AB_2095847), phospho-PERK (rabbit, 1:1000, Cell Signaling Technology, Cat# 3179S, RRID: AB_2095853), eukaryotic translation initiation factor 2 subunit alpha (eIF2α; rabbit, 1:1000, Cell Signaling Technology, Cat# 9722S, RRID: AB_2230924), phospho-eIF2α (rabbit, 1:1000, Cell Signaling Technology, 9721S, RRID: AB_330951), phosphatidylinositol 3-kinase regulatory subunit alpha (PI3 kinase p85; rabbit, 1:1000, Cell Signaling Technology, Cat# 4257S, RRID: AB_659889), RAC-alpha serine/threonine-protein kinase (AKT; rabbit, 1:1000, Cell Signaling Technology, Cat# 4691S, RRID: AB_915783), phospho-AKT (rabbit, 1:1000, Cell Signaling Technology, Cat# 4060S, RRID: AB_2315049), mammalian target of rapamycin (mTOR; rabbit, 1:1000, Cell Signaling Technology, Cat# 2983S, RRID: AB_2105622), phospho-mTOR (rabbit, 1:1000, Cell Signaling Technology, Cat# 2971S, RRID: AB_330970), and β-actin (mouse, 1:1000, Proteintech, Cat# 66009-1-Ig, RRID: AB_2687938).

    Techniques: Modification, Staining, TUNEL Assay, Recombinant

    rtPA attenuates autophagy in peri-hematoma tissue after ICH. (A–D) Analysis of autophagy-associated proteins of peri-hematoma tissue at 24 hours after treatment. (E–H) Analysis of autophagy-associated proteins of peri-hematoma tissue at 72 hours after treatment. Data are shown as mean ± SEM ( n = 3 per group). ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . sham group; & P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, #### P < 0.0001, vs . ICH + vehicle group (one-way analysis of variance followed by Tukey’s post hoc test). beclin1: Coiled-coil myosin-like bcl2-interacting protein; ICH: intracerebral hemorrhage; LC3: microtubule-associated proteins 1A/1B light chain 3B; p62: sequestosome-1/ubiquitin-binding protein p62; rtPA: recombinant tissue plasminogen activator.

    Journal: Neural Regeneration Research

    Article Title: Recombinant tissue plasminogen activator protects neurons after intracerebral hemorrhage through activating the PI3K/AKT/mTOR pathway

    doi: 10.4103/NRR.NRR-D-23-01953

    Figure Lengend Snippet: rtPA attenuates autophagy in peri-hematoma tissue after ICH. (A–D) Analysis of autophagy-associated proteins of peri-hematoma tissue at 24 hours after treatment. (E–H) Analysis of autophagy-associated proteins of peri-hematoma tissue at 72 hours after treatment. Data are shown as mean ± SEM ( n = 3 per group). ** P < 0.01, *** P < 0.001, **** P < 0.0001, vs . sham group; & P < 0.05, && P < 0.01, &&&& P < 0.0001, vs . ICH group; # P < 0.05, ## P < 0.01, #### P < 0.0001, vs . ICH + vehicle group (one-way analysis of variance followed by Tukey’s post hoc test). beclin1: Coiled-coil myosin-like bcl2-interacting protein; ICH: intracerebral hemorrhage; LC3: microtubule-associated proteins 1A/1B light chain 3B; p62: sequestosome-1/ubiquitin-binding protein p62; rtPA: recombinant tissue plasminogen activator.

    Article Snippet: The following primary antibodies were used for analysis: bcl2 (rabbit, 1:1000, Proteintech, Cat# 12789-1-AP, RRID: AB_2227948), bax (rabbit, 1:1000, Proteintech, Cat# 50599-2-Ig, RRID: AB_2061561), coiled-coil myosin-like bcl2-interacting protein (beclin1; rabbit, 1:1000, Proteintech, Cat# 11306-1-AP, RRID: AB_2259061), sequestosome-1/ubiquitin-binding protein p62 (SQSTM1/p62; rabbit, 1:1000, Abclonal, Cat# A11250, RRID: AB_2758477), microtubule-associated proteins 1A/1B light chain 3B (LC3; rabbit, 1:1000, Abcam, Cat# ab48394, RRID: AB_881433), endoplasmic reticulum chaperone BiP (Grp78/BIP; mouse, 1:1000, Proteintech, Cat# 66574-1-Ig, RRID: AB_2881934), cyclic AMP-dependent transcription factor ATF-6 alpha (ATF6; rabbit, 1:1000, Proteintech, Cat# 24169-1-AP, RRID: AB_2876891), PRKR-like endoplasmic reticulum kinase (PERK; rabbit, 1:1000, Cell Signaling Technology, Cat# 3192S, RRID: AB_2095847), phospho-PERK (rabbit, 1:1000, Cell Signaling Technology, Cat# 3179S, RRID: AB_2095853), eukaryotic translation initiation factor 2 subunit alpha (eIF2α; rabbit, 1:1000, Cell Signaling Technology, Cat# 9722S, RRID: AB_2230924), phospho-eIF2α (rabbit, 1:1000, Cell Signaling Technology, 9721S, RRID: AB_330951), phosphatidylinositol 3-kinase regulatory subunit alpha (PI3 kinase p85; rabbit, 1:1000, Cell Signaling Technology, Cat# 4257S, RRID: AB_659889), RAC-alpha serine/threonine-protein kinase (AKT; rabbit, 1:1000, Cell Signaling Technology, Cat# 4691S, RRID: AB_915783), phospho-AKT (rabbit, 1:1000, Cell Signaling Technology, Cat# 4060S, RRID: AB_2315049), mammalian target of rapamycin (mTOR; rabbit, 1:1000, Cell Signaling Technology, Cat# 2983S, RRID: AB_2105622), phospho-mTOR (rabbit, 1:1000, Cell Signaling Technology, Cat# 2971S, RRID: AB_330970), and β-actin (mouse, 1:1000, Proteintech, Cat# 66009-1-Ig, RRID: AB_2687938).

    Techniques: Ubiquitin Proteomics, Binding Assay, Recombinant

    rtPA attenuates neuron apoptosis and autophagy after experimental ICH in vitro . (A–C) The DEGs between control group and hemin group associated with autophagy animals (KEGG: mmu04140), positive regulation of neuron apoptotic process (GO: 0043525), and positive regulation of response to endoplasmic reticulum stress (GO: 1905898) were screened, and the transcriptional levels of DEGs in each group are presented as heatmaps. (D, E) Analysis of apoptosis-associated proteins. (F) Transmission electron microscopy images of neurons after hemin and rtPA treatment. Red asterisk indicates the autophagosome, black arrows indicate the endoplasmic reticulum, and N means nucleus. Scale bars: 1 µm. (G–J) Analysis of autophagy-associated proteins. Data are shown as mean ± SEM ( n = 3–4). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs. hemin group (one-way analysis of variance followed by Tukey’s post hoc test). bax: Apoptosis regulator bax; bcl2: apoptosis regulator bcl2; beclin1: coiled-coil myosin-like bcl2-interacting protein; DEGs: differential expression genes; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; LC3: microtubule-associated proteins 1A/1B light chain 3B; p62: sequestosome-1/ubiquitin-binding protein p62; rtPA: recombinant tissue plasminogen activator.

    Journal: Neural Regeneration Research

    Article Title: Recombinant tissue plasminogen activator protects neurons after intracerebral hemorrhage through activating the PI3K/AKT/mTOR pathway

    doi: 10.4103/NRR.NRR-D-23-01953

    Figure Lengend Snippet: rtPA attenuates neuron apoptosis and autophagy after experimental ICH in vitro . (A–C) The DEGs between control group and hemin group associated with autophagy animals (KEGG: mmu04140), positive regulation of neuron apoptotic process (GO: 0043525), and positive regulation of response to endoplasmic reticulum stress (GO: 1905898) were screened, and the transcriptional levels of DEGs in each group are presented as heatmaps. (D, E) Analysis of apoptosis-associated proteins. (F) Transmission electron microscopy images of neurons after hemin and rtPA treatment. Red asterisk indicates the autophagosome, black arrows indicate the endoplasmic reticulum, and N means nucleus. Scale bars: 1 µm. (G–J) Analysis of autophagy-associated proteins. Data are shown as mean ± SEM ( n = 3–4). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . control group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs. hemin group (one-way analysis of variance followed by Tukey’s post hoc test). bax: Apoptosis regulator bax; bcl2: apoptosis regulator bcl2; beclin1: coiled-coil myosin-like bcl2-interacting protein; DEGs: differential expression genes; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; LC3: microtubule-associated proteins 1A/1B light chain 3B; p62: sequestosome-1/ubiquitin-binding protein p62; rtPA: recombinant tissue plasminogen activator.

    Article Snippet: The following primary antibodies were used for analysis: bcl2 (rabbit, 1:1000, Proteintech, Cat# 12789-1-AP, RRID: AB_2227948), bax (rabbit, 1:1000, Proteintech, Cat# 50599-2-Ig, RRID: AB_2061561), coiled-coil myosin-like bcl2-interacting protein (beclin1; rabbit, 1:1000, Proteintech, Cat# 11306-1-AP, RRID: AB_2259061), sequestosome-1/ubiquitin-binding protein p62 (SQSTM1/p62; rabbit, 1:1000, Abclonal, Cat# A11250, RRID: AB_2758477), microtubule-associated proteins 1A/1B light chain 3B (LC3; rabbit, 1:1000, Abcam, Cat# ab48394, RRID: AB_881433), endoplasmic reticulum chaperone BiP (Grp78/BIP; mouse, 1:1000, Proteintech, Cat# 66574-1-Ig, RRID: AB_2881934), cyclic AMP-dependent transcription factor ATF-6 alpha (ATF6; rabbit, 1:1000, Proteintech, Cat# 24169-1-AP, RRID: AB_2876891), PRKR-like endoplasmic reticulum kinase (PERK; rabbit, 1:1000, Cell Signaling Technology, Cat# 3192S, RRID: AB_2095847), phospho-PERK (rabbit, 1:1000, Cell Signaling Technology, Cat# 3179S, RRID: AB_2095853), eukaryotic translation initiation factor 2 subunit alpha (eIF2α; rabbit, 1:1000, Cell Signaling Technology, Cat# 9722S, RRID: AB_2230924), phospho-eIF2α (rabbit, 1:1000, Cell Signaling Technology, 9721S, RRID: AB_330951), phosphatidylinositol 3-kinase regulatory subunit alpha (PI3 kinase p85; rabbit, 1:1000, Cell Signaling Technology, Cat# 4257S, RRID: AB_659889), RAC-alpha serine/threonine-protein kinase (AKT; rabbit, 1:1000, Cell Signaling Technology, Cat# 4691S, RRID: AB_915783), phospho-AKT (rabbit, 1:1000, Cell Signaling Technology, Cat# 4060S, RRID: AB_2315049), mammalian target of rapamycin (mTOR; rabbit, 1:1000, Cell Signaling Technology, Cat# 2983S, RRID: AB_2105622), phospho-mTOR (rabbit, 1:1000, Cell Signaling Technology, Cat# 2971S, RRID: AB_330970), and β-actin (mouse, 1:1000, Proteintech, Cat# 66009-1-Ig, RRID: AB_2687938).

    Techniques: In Vitro, Control, Transmission Assay, Electron Microscopy, Quantitative Proteomics, Ubiquitin Proteomics, Binding Assay, Recombinant