Journal: BMC Molecular and Cell Biology

Article Title: Alcohol exposure induces ferroptosis-dominated programmed cell death in esophageal epithelial cells

doi: 10.1186/s12860-026-00589-5

Figure Lengend Snippet: A short-term exposure to 5.0% ethanol triggers autophagy that supports cell growth. A . An equal number of HEEC cells was seeded on coverslips and incubated with complete medium (control) or medium containing 5.0% ethanol for 5 min, 30 min, or 2, 6, and 12 h. Cells were fixed in 4% paraformaldehyde and stained for LC3B. An FITC-conjugated secondary antibody was used to develop the signal. Cell nuclei were counterstained with DAPI. Scale bar = 5 μm. B . An equal number of HEEC cells was seeded in 96-well plates and incubated with complete medium (control), medium containing 5.0% ethanol, or medium containing 5.0% ethanol plus 10 µM Chloroquine (CQ) for 5 min, 30 min, 2 h, 6 h, or 12 h. Cell viability was assessed using CCK-8 kits. * indicates a significant change between EtOH and EtOH/CQ. C . An equal number of Het1A cells was seeded in 96-well plates and incubated with the complete medium containing DMSO (control), the medium containing 5.0% ethanol, or the medium containing 5.0% ethanol plus 10 µM Chloroquine (CQ) for 5 min, 30 min, 2 h, 6 h, or 12 h. Cell viability was assessed using CCK-8 kits. * indicates a significant change between EtOH and EtOH/CQ. D . An equal number of HEEC or Het1A cells was seeded on the coverslips and incubated with the complete medium containing DMSO (control), the medium containing 5.0% ethanol, or the medium containing 5.0% ethanol plus 10 µM Chloroquine (CQ) for 30 min. Cells were fixed in 4% paraformaldehyde and stained for PCNA. An FITC-conjugated secondary antibody was used to develop the signal. Cell nuclei were counterstained with DAPI. Scale bar = 5 μm. E . An equal number of HEEC or Het1A cells was seeded in 6-well plates and incubated with complete medium containing DMSO (control), medium containing 5.0% ethanol, or medium containing 5.0% ethanol plus 10 µM Chloroquine (CQ) for 30 min. Protein was extracted and analyzed by Western blotting. The membranes were probed for LC3B, PCNA, Beclin-1, or GAPDH. F . Quantitative analyses of LC3B, PCNA, and Beclin-1 expression against GAPDH in HEEC cells. * indicates a significant change compared to the control (DMSO). G . Quantitative analyses of LC3B, PCNA, and Beclin-1 expression against GAPDH in Het1A cells. * indicates a significant change compared to the control (DMSO)

Article Snippet: The following primary antibodies were used: Beclin-1 (Origene, #TA502643), PCNA, CASP1 (Abcam, #ab179515), CASP3 (Santa Cruz Biotechnology, #sc-271759), CASP8 (Origene, #TA374288), CASP9 (Origene, #TA4227045), endoG, GPX4 (Origene, #TA423164M), GSDMD, IL-1β (Santa Cruz Biotechnology, #sc-12742), MLKL, phosphor-MLKL (Abcam, #ab196436), SLC7A11 (Origene, #TA423232), LC3B, BAX, GAPDH (OriGene, #TA800894), and β-actin (Santa Cruz Biotechnology, Inc. #sc-69879).

Techniques: Incubation, Control, Staining, CCK-8 Assay, Western Blot, Expressing

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

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

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

Journal: Chinese Herbal Medicines

Article Title: Isoquinoline alkaloids in Coptis chinensis to treat Alzheimer’s disease through promoting growth of Bifidobacterium breve inhibiting abnormal autophagy using a novel AI high-content intelligent imaging system

doi: 10.1016/j.chmed.2026.02.003

Figure Lengend Snippet: Co-administration of C. chinensis and B. breve S1 treated AD mice by regulating autophagy. (A, D) Expression of p62 protein ( n = 5). (B, E) Expression of Beclin-1 protein ( n = 5). (C, F) Expression of LC3I and LC3II protein ( n = 5). (G) Determination of content of BDNF ( n = 5). (H) Determination of content of NGF ( n = 5). (I) Determination of content of AGEs ( n = 5). (J) Determination of content of sRAGE ( n = 5). n. s., no significant difference, ### P < 0.001 vs Control group, * P < 0.05 ** P < 0.01 *** P < 0.001 vs Model-free group.

Article Snippet: Antibody to microtubule-associated protein 1 light chain 3 (LC3, lot number: 4), antibody to Sequestosome 1 (SQSTM1/p62, lot number: 6), antibody to Beclin-1 (Beclin-1/BECN1, lot number: 5) were purchased from Cell Signaling Technology (Shanghai, China).

Techniques: Expressing, Control

Journal: Chinese Herbal Medicines

Article Title: Isoquinoline alkaloids in Coptis chinensis to treat Alzheimer’s disease through promoting growth of Bifidobacterium breve inhibiting abnormal autophagy using a novel AI high-content intelligent imaging system

doi: 10.1016/j.chmed.2026.02.003

Figure Lengend Snippet: The expression difference of LC3II , p62 , Beclin-1 genes in different cell clusters ( brain-map.org ). (A) The expression difference of LC3Ⅱ. (B) The expression difference of p62. (C) The expression difference of Beclin-1.

Article Snippet: Antibody to microtubule-associated protein 1 light chain 3 (LC3, lot number: 4), antibody to Sequestosome 1 (SQSTM1/p62, lot number: 6), antibody to Beclin-1 (Beclin-1/BECN1, lot number: 5) were purchased from Cell Signaling Technology (Shanghai, China).

Techniques: Expressing

Journal: Chinese Herbal Medicines

Article Title: Isoquinoline alkaloids in Coptis chinensis to treat Alzheimer’s disease through promoting growth of Bifidobacterium breve inhibiting abnormal autophagy using a novel AI high-content intelligent imaging system

doi: 10.1016/j.chmed.2026.02.003

Figure Lengend Snippet: Magnoflorine, palmatrubine, 13-methylberberine regulate autophagy in cells. (A, B) Expression of p62 protein ( n = 5). (C, D) Expression of Beclin-1 protein ( n = 5). (E, F) Expression of LC3I and LC3II protein ( n = 5). (G) Determination of content of BNDF ( n = 5). (H) Determination of content of GDNF ( n = 5). (I) Determination of content of NGF ( n = 5). (J) Determination of content of ROS ( n = 5). n. s., no significant difference, ### P < 0.001 vs Control group, * P < 0.05 ** P < 0.01 *** P < 0.001 vs Model group.

Article Snippet: Antibody to microtubule-associated protein 1 light chain 3 (LC3, lot number: 4), antibody to Sequestosome 1 (SQSTM1/p62, lot number: 6), antibody to Beclin-1 (Beclin-1/BECN1, lot number: 5) were purchased from Cell Signaling Technology (Shanghai, China).

Techniques: Expressing, Control