Review

grp78 (Proteintech)

Proteintech is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Proteintech grp78

Interaction between PRAS40 and <t>GRP78.</t> (A) Prediction of PRAS40’s binding proteins involved in ER stress and UPR by overlapping the PRAS40-binding proteins determined by Co-IP-MS and the ER stress- and UPR-related factors overexpressed in TCGA-CRC samples. (B) The unique peptides of GRP78 enriched in PRAS40-bound precipitates were determined by MS. (C-F) Co-IP analyses in HEK-293T and HT29 cells transfected with empty vector or Flag-PRAS40 expression vector (C, E), Flag-GRP78 expression vector (D, F). (G) GST pull-down assays. (H) Immunofluorescence staining with anti-PRAS40 and anti-GRP78 antibodies in HCT116 cells. (I) Co-IP analyses in HEK-293T cells transfected with empty vector or expression vectors of Flag-PRAS40 deletion mutants. Scale bar, 10 μm.

Grp78, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/anti+bip/pmc13020080-49-9-18?v=Proteintech
Average 95 stars, based on 33 article reviews

grp78 - by Bioz Stars, 2026-07

95/100 stars

Images

1) Product Images from "PRAS40 activates the IRE1α-XBP-1-mediated unfolded protein response to exacerbate colorectal cancer by enhancing ST6Gal1-dependent α-2, 6 sialylation of GRP78"

Article Title: PRAS40 activates the IRE1α-XBP-1-mediated unfolded protein response to exacerbate colorectal cancer by enhancing ST6Gal1-dependent α-2, 6 sialylation of GRP78

Journal: Neoplasia (New York, N.Y.)

doi: 10.1016/j.neo.2026.101297

Interaction between PRAS40 and GRP78. (A) Prediction of PRAS40’s binding proteins involved in ER stress and UPR by overlapping the PRAS40-binding proteins determined by Co-IP-MS and the ER stress- and UPR-related factors overexpressed in TCGA-CRC samples. (B) The unique peptides of GRP78 enriched in PRAS40-bound precipitates were determined by MS. (C-F) Co-IP analyses in HEK-293T and HT29 cells transfected with empty vector or Flag-PRAS40 expression vector (C, E), Flag-GRP78 expression vector (D, F). (G) GST pull-down assays. (H) Immunofluorescence staining with anti-PRAS40 and anti-GRP78 antibodies in HCT116 cells. (I) Co-IP analyses in HEK-293T cells transfected with empty vector or expression vectors of Flag-PRAS40 deletion mutants. Scale bar, 10 μm.

Figure Legend Snippet: Interaction between PRAS40 and GRP78. (A) Prediction of PRAS40’s binding proteins involved in ER stress and UPR by overlapping the PRAS40-binding proteins determined by Co-IP-MS and the ER stress- and UPR-related factors overexpressed in TCGA-CRC samples. (B) The unique peptides of GRP78 enriched in PRAS40-bound precipitates were determined by MS. (C-F) Co-IP analyses in HEK-293T and HT29 cells transfected with empty vector or Flag-PRAS40 expression vector (C, E), Flag-GRP78 expression vector (D, F). (G) GST pull-down assays. (H) Immunofluorescence staining with anti-PRAS40 and anti-GRP78 antibodies in HCT116 cells. (I) Co-IP analyses in HEK-293T cells transfected with empty vector or expression vectors of Flag-PRAS40 deletion mutants. Scale bar, 10 μm.

Techniques Used: Binding Assay, Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation, Expressing, Immunofluorescence, Staining

Effects of PRAS40 on the N-glycosylation of GRP78. (A) ssGSEA for the correlation between PRAS40 and N-glycan biosynthesis in TCGA-CRC samples. (B) SNA blotting analyses of peri-cancer and cancer tissues from mouse CRC models. (C-D) SNA blotting analyses of the HCT116 cells transfected with empty vector or Flag-PRAS40 expression vector (C) and control or PRAS40 shRNA (D). (E-G) SNA-pull down-MS in the cells transfected with empty vector or Flag-PRAS40 expression vector. CBB staining (E), the unique peptides spectrum of GRP78 (F) and the LC-MS/MS of amino acid 50-60 of GRP78 (G). (H) The HCT116 cells transfected with empty vector or Flag-PRAS40 expression vector was treated with or without PNGase F (1 mU), followed by western blotting analyses. (I-J) The HCT116 cells transfected with empty vector or Flag-PRAS40 expression vector (I), or control or PRAS40 shRNA, followed by SNA blotting analyses (J).

Figure Legend Snippet: Effects of PRAS40 on the N-glycosylation of GRP78. (A) ssGSEA for the correlation between PRAS40 and N-glycan biosynthesis in TCGA-CRC samples. (B) SNA blotting analyses of peri-cancer and cancer tissues from mouse CRC models. (C-D) SNA blotting analyses of the HCT116 cells transfected with empty vector or Flag-PRAS40 expression vector (C) and control or PRAS40 shRNA (D). (E-G) SNA-pull down-MS in the cells transfected with empty vector or Flag-PRAS40 expression vector. CBB staining (E), the unique peptides spectrum of GRP78 (F) and the LC-MS/MS of amino acid 50-60 of GRP78 (G). (H) The HCT116 cells transfected with empty vector or Flag-PRAS40 expression vector was treated with or without PNGase F (1 mU), followed by western blotting analyses. (I-J) The HCT116 cells transfected with empty vector or Flag-PRAS40 expression vector (I), or control or PRAS40 shRNA, followed by SNA blotting analyses (J).

Techniques Used: Glycoproteomics, Transfection, Plasmid Preparation, Expressing, Control, shRNA, Staining, Liquid Chromatography with Mass Spectroscopy, Western Blot

Effects of the N-glycosylation at Asn59 on the function of GRP78 and the UPR. (A) Predicted N-glycosylation sites in GRP78 by NetNGlyc-1.0 ( https://services.healthtech.dtu.dk/services/NetNGlyc-1.0/ ). (B) Molecular docking of the interaction between PRAS40 and GRP78. PRAS40 was shown in red, and GRP78 was shown in blue, respectively. Hydrogen bonds were indicated by yellow dashed lines, and the numbers represented the lengths of hydrogen bonds. (C) The solvation energy effects (∆iG) of the interaction between PRAS40 and the predicted residues of GRP78 ( https://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ). (D) The cells deleted with GRP78 were overexpressed with shRNA-resistant wild type GRP78 or GRP78 N59Q , followed with SNA pull down assays. (E-J) The cells deleted with GRP78 was overexpressed with Flag-PRAS40 together with or without shRNA-resistant wild type GRP78 or GRP78 N59Q , were treated with or without Tg. Cell viability analyses (E-F), flow cytometry analyses and the quantification of 3 experiments (G-H), western blotting analyses (I) and PCR analysis (J).

Figure Legend Snippet: Effects of the N-glycosylation at Asn59 on the function of GRP78 and the UPR. (A) Predicted N-glycosylation sites in GRP78 by NetNGlyc-1.0 ( https://services.healthtech.dtu.dk/services/NetNGlyc-1.0/ ). (B) Molecular docking of the interaction between PRAS40 and GRP78. PRAS40 was shown in red, and GRP78 was shown in blue, respectively. Hydrogen bonds were indicated by yellow dashed lines, and the numbers represented the lengths of hydrogen bonds. (C) The solvation energy effects (∆iG) of the interaction between PRAS40 and the predicted residues of GRP78 ( https://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ). (D) The cells deleted with GRP78 were overexpressed with shRNA-resistant wild type GRP78 or GRP78 N59Q , followed with SNA pull down assays. (E-J) The cells deleted with GRP78 was overexpressed with Flag-PRAS40 together with or without shRNA-resistant wild type GRP78 or GRP78 N59Q , were treated with or without Tg. Cell viability analyses (E-F), flow cytometry analyses and the quantification of 3 experiments (G-H), western blotting analyses (I) and PCR analysis (J).

Techniques Used: Glycoproteomics, shRNA, Flow Cytometry, Western Blot

Effects of ST6Gal1-dependent α-2, 6 sialylation of GRP78 on the UPR. (A) Molecular docking of the interaction between GRP78 and ST6Gal1. GRP78 was shown in blue, and ST6Gal1 was shown in green, respectively. Hydrogen bonds were indicated by yellow dashed lines, and the numbers represented the lengths of hydrogen bonds. (B) Immunofluorescence staining with anti-GRP78 and anti-ST6Gal1 antibodies in HCT116 cells. (C-D) Co-IP followed by western blotting analyses in PRAS40-overexpressed HCT116 cells with anti-ST6Gal1 (C) and anti-GRP78 antibodies (D), respectively. (E-K) The HCT116 cells deleted with ST6Gal1 and overexpressed with Flag-PRAS40, were treated with or without Tg. SNA-pull down assays (E), cell viability analyses (F-G), flow cytometry analyses and the quantification of 3 experiments (H-I), western blotting analyses (J) and PCR analysis (K). Data represent the mean ± SD. Scale bar, 10 μm. ** P < 0.01; *** P < 0.001.

Figure Legend Snippet: Effects of ST6Gal1-dependent α-2, 6 sialylation of GRP78 on the UPR. (A) Molecular docking of the interaction between GRP78 and ST6Gal1. GRP78 was shown in blue, and ST6Gal1 was shown in green, respectively. Hydrogen bonds were indicated by yellow dashed lines, and the numbers represented the lengths of hydrogen bonds. (B) Immunofluorescence staining with anti-GRP78 and anti-ST6Gal1 antibodies in HCT116 cells. (C-D) Co-IP followed by western blotting analyses in PRAS40-overexpressed HCT116 cells with anti-ST6Gal1 (C) and anti-GRP78 antibodies (D), respectively. (E-K) The HCT116 cells deleted with ST6Gal1 and overexpressed with Flag-PRAS40, were treated with or without Tg. SNA-pull down assays (E), cell viability analyses (F-G), flow cytometry analyses and the quantification of 3 experiments (H-I), western blotting analyses (J) and PCR analysis (K). Data represent the mean ± SD. Scale bar, 10 μm. ** P < 0.01; *** P < 0.001.

Techniques Used: Immunofluorescence, Staining, Co-Immunoprecipitation Assay, Western Blot, Flow Cytometry

Image Search Results

Journal: Neoplasia (New York, N.Y.)

Article Title: PRAS40 activates the IRE1α-XBP-1-mediated unfolded protein response to exacerbate colorectal cancer by enhancing ST6Gal1-dependent α-2, 6 sialylation of GRP78

doi: 10.1016/j.neo.2026.101297

Figure Lengend Snippet: Interaction between PRAS40 and GRP78. (A) Prediction of PRAS40’s binding proteins involved in ER stress and UPR by overlapping the PRAS40-binding proteins determined by Co-IP-MS and the ER stress- and UPR-related factors overexpressed in TCGA-CRC samples. (B) The unique peptides of GRP78 enriched in PRAS40-bound precipitates were determined by MS. (C-F) Co-IP analyses in HEK-293T and HT29 cells transfected with empty vector or Flag-PRAS40 expression vector (C, E), Flag-GRP78 expression vector (D, F). (G) GST pull-down assays. (H) Immunofluorescence staining with anti-PRAS40 and anti-GRP78 antibodies in HCT116 cells. (I) Co-IP analyses in HEK-293T cells transfected with empty vector or expression vectors of Flag-PRAS40 deletion mutants. Scale bar, 10 μm.

Article Snippet: Antibodies were purchased for detection of PRAS40 (Cell Signaling); GRP78, XBP-1, PARP, IRE1, Flag, GST, ST6Gal1, α-tubulin, β-actin (Proteintech); and SNA (Vector Laboratories).

Techniques: Binding Assay, Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation, Expressing, Immunofluorescence, Staining

Journal: Neoplasia (New York, N.Y.)

Article Title: PRAS40 activates the IRE1α-XBP-1-mediated unfolded protein response to exacerbate colorectal cancer by enhancing ST6Gal1-dependent α-2, 6 sialylation of GRP78

doi: 10.1016/j.neo.2026.101297

Figure Lengend Snippet: Effects of PRAS40 on the N-glycosylation of GRP78. (A) ssGSEA for the correlation between PRAS40 and N-glycan biosynthesis in TCGA-CRC samples. (B) SNA blotting analyses of peri-cancer and cancer tissues from mouse CRC models. (C-D) SNA blotting analyses of the HCT116 cells transfected with empty vector or Flag-PRAS40 expression vector (C) and control or PRAS40 shRNA (D). (E-G) SNA-pull down-MS in the cells transfected with empty vector or Flag-PRAS40 expression vector. CBB staining (E), the unique peptides spectrum of GRP78 (F) and the LC-MS/MS of amino acid 50-60 of GRP78 (G). (H) The HCT116 cells transfected with empty vector or Flag-PRAS40 expression vector was treated with or without PNGase F (1 mU), followed by western blotting analyses. (I-J) The HCT116 cells transfected with empty vector or Flag-PRAS40 expression vector (I), or control or PRAS40 shRNA, followed by SNA blotting analyses (J).

Techniques: Glycoproteomics, Transfection, Plasmid Preparation, Expressing, Control, shRNA, Staining, Liquid Chromatography with Mass Spectroscopy, Western Blot

Journal: Neoplasia (New York, N.Y.)

Article Title: PRAS40 activates the IRE1α-XBP-1-mediated unfolded protein response to exacerbate colorectal cancer by enhancing ST6Gal1-dependent α-2, 6 sialylation of GRP78

doi: 10.1016/j.neo.2026.101297

Figure Lengend Snippet: Effects of the N-glycosylation at Asn59 on the function of GRP78 and the UPR. (A) Predicted N-glycosylation sites in GRP78 by NetNGlyc-1.0 ( https://services.healthtech.dtu.dk/services/NetNGlyc-1.0/ ). (B) Molecular docking of the interaction between PRAS40 and GRP78. PRAS40 was shown in red, and GRP78 was shown in blue, respectively. Hydrogen bonds were indicated by yellow dashed lines, and the numbers represented the lengths of hydrogen bonds. (C) The solvation energy effects (∆iG) of the interaction between PRAS40 and the predicted residues of GRP78 ( https://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ). (D) The cells deleted with GRP78 were overexpressed with shRNA-resistant wild type GRP78 or GRP78 N59Q , followed with SNA pull down assays. (E-J) The cells deleted with GRP78 was overexpressed with Flag-PRAS40 together with or without shRNA-resistant wild type GRP78 or GRP78 N59Q , were treated with or without Tg. Cell viability analyses (E-F), flow cytometry analyses and the quantification of 3 experiments (G-H), western blotting analyses (I) and PCR analysis (J).

Techniques: Glycoproteomics, shRNA, Flow Cytometry, Western Blot

Journal: Neoplasia (New York, N.Y.)

Article Title: PRAS40 activates the IRE1α-XBP-1-mediated unfolded protein response to exacerbate colorectal cancer by enhancing ST6Gal1-dependent α-2, 6 sialylation of GRP78

doi: 10.1016/j.neo.2026.101297

Figure Lengend Snippet: Effects of ST6Gal1-dependent α-2, 6 sialylation of GRP78 on the UPR. (A) Molecular docking of the interaction between GRP78 and ST6Gal1. GRP78 was shown in blue, and ST6Gal1 was shown in green, respectively. Hydrogen bonds were indicated by yellow dashed lines, and the numbers represented the lengths of hydrogen bonds. (B) Immunofluorescence staining with anti-GRP78 and anti-ST6Gal1 antibodies in HCT116 cells. (C-D) Co-IP followed by western blotting analyses in PRAS40-overexpressed HCT116 cells with anti-ST6Gal1 (C) and anti-GRP78 antibodies (D), respectively. (E-K) The HCT116 cells deleted with ST6Gal1 and overexpressed with Flag-PRAS40, were treated with or without Tg. SNA-pull down assays (E), cell viability analyses (F-G), flow cytometry analyses and the quantification of 3 experiments (H-I), western blotting analyses (J) and PCR analysis (K). Data represent the mean ± SD. Scale bar, 10 μm. ** P < 0.01; *** P < 0.001.

Techniques: Immunofluorescence, Staining, Co-Immunoprecipitation Assay, Western Blot, Flow Cytometry

TNKS affects distributions of Formin2 and ER for chromosome migration in mouse oocytes. (A) Representative images of Formin2 in MI stage oocyte from the control and JW55 treatment groups. Green, Formin2. Cyan, DNA. Bar = 20 μm. (B) Relative intensity of Formin2 at spindle poles in oocytes from the control (n = 47) and JW55 treatment (n = 51) groups. ***, P < 0.001. (C) Band intensity analysis of Formin2 expression in the MI stage oocytes from control and JW55 treatment groups. *, P < 0.05. (D) Representative images of ER-tracker in MI stage oocyte from the control and JW55 treatment groups. Black, ER-tracker. Bar = 20 μm. (E) The percentage of abnormal ER distribution in oocytes from the control (n = 50) and JW55-treated (n = 56) groups. *, P < 0.05. (F) Relative intensity of ER-tracker in oocytes from the control (n = 50) and JW55-treated (n = 56) groups. ***, P < 0.001. (G) Western blot of GRP78 in MI stage oocytes from control and JW55 treatment groups. (H) Band intensity analysis of GRP78 in the control and JW55 treatment groups. *, P < 0.05. (I) Representative images of a relative position change between chromosomes and Formin2 in ATI stage oocytes from the control and JW55 treatment groups. Green, Formin2. Cyan, DNA. Bar = 20 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Journal: Journal of Advanced Research

Article Title: Tankyrase activity is essential for asymmetric division and chromosome segregation in oocyte meiosis

doi: 10.1016/j.jare.2025.07.008

Figure Lengend Snippet: TNKS affects distributions of Formin2 and ER for chromosome migration in mouse oocytes. (A) Representative images of Formin2 in MI stage oocyte from the control and JW55 treatment groups. Green, Formin2. Cyan, DNA. Bar = 20 μm. (B) Relative intensity of Formin2 at spindle poles in oocytes from the control (n = 47) and JW55 treatment (n = 51) groups. ***, P < 0.001. (C) Band intensity analysis of Formin2 expression in the MI stage oocytes from control and JW55 treatment groups. *, P < 0.05. (D) Representative images of ER-tracker in MI stage oocyte from the control and JW55 treatment groups. Black, ER-tracker. Bar = 20 μm. (E) The percentage of abnormal ER distribution in oocytes from the control (n = 50) and JW55-treated (n = 56) groups. *, P < 0.05. (F) Relative intensity of ER-tracker in oocytes from the control (n = 50) and JW55-treated (n = 56) groups. ***, P < 0.001. (G) Western blot of GRP78 in MI stage oocytes from control and JW55 treatment groups. (H) Band intensity analysis of GRP78 in the control and JW55 treatment groups. *, P < 0.05. (I) Representative images of a relative position change between chromosomes and Formin2 in ATI stage oocytes from the control and JW55 treatment groups. Green, Formin2. Cyan, DNA. Bar = 20 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Rabbit anti-α-Tubulin antibody (11224-1-AP), rabbit polyclonal anti-GAPDH antibody (10494-1-AP), rabbit anti-GRP78 antibody (11587-1-AP), rabbit anti-TNKS1 antibody (18030-1-AP), rabbit anti-Ran antibody (10469-1-AP), and rabbit anti-Rab11a antibody (67902-1-Ig), rabbit anti-Securin antibody (18040-1-AP), rabbit anti-CDC25C antibody (66912-1-Ig), rabbit anti-Formin2 antibody (11259-1-AP), rabbit anti-BubR1 (11504-2-AP) were from Proteintech.

Techniques: Migration, Control, Expressing, Western Blot

Experiment design. (A) In Experiment 1, the effect of rtPA on the ICH mouse model was investigated. (B) In Experiment 2, the effect and the possible mechanisms of rtPA in the ICH model using primary cortical neurons in vitro were investigated. (C) In Experiment 3, the mechanism of rtPA’s effect on the ICH model in neurons in vitro was examined using the PI3K pathway inhibitor. (D) In Experiment 4, the protein domain that mediates rtPA’s neuroprotective effect in the ICH model in neurons in vitro was investigated. DMSO: Dimethyl sulfoxide; 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: Experiment design. (A) In Experiment 1, the effect of rtPA on the ICH mouse model was investigated. (B) In Experiment 2, the effect and the possible mechanisms of rtPA in the ICH model using primary cortical neurons in vitro were investigated. (C) In Experiment 3, the mechanism of rtPA’s effect on the ICH model in neurons in vitro was examined using the PI3K pathway inhibitor. (D) In Experiment 4, the protein domain that mediates rtPA’s neuroprotective effect in the ICH model in neurons in vitro was investigated. DMSO: Dimethyl sulfoxide; 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: In Vitro, Staining, Recombinant, TUNEL Assay

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.

Techniques: Modification, Staining, TUNEL 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.

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

rtPA ameliorates endoplasmic reticulum stress in the in vitro ICH cell model. (A) Confocal images and three-dimensional reconstruction of endoplasmic reticulum continuity of neurons by ER tracker after hemin and rtPA treatment. Scale bars: 3 µm. (B–F) Quantitative analysis of ER stress–associated proteins of neurons. Data are shown as mean ± SEM ( n = 3 per group). * P < 0.05, ** P < 0.01, vs. control group; # P < 0.05, ## P < 0.01, vs . hemin group (one-way analysis of variance followed by Tukey’s post hoc test). (G) Immunofluorescence staining images of p-PERK (red, labeled by Cy3) in neurons after hemin and rtPA treatment. Scale bars: 50 µm. 3D: Three-dimensional; ATF6: cyclic AMP-dependent transcription factor ATF-6 alpha; DAPI: 4′,6-diamidino-2-phenylindole; eIF2α: eukaryotic translation initiation factor 2 subunit alpha; ER: endoplasmic reticulum; Grp78: endoplasmic reticulum chaperone BiP; PERK: PRKR-like endoplasmic reticulum kinase; 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 ameliorates endoplasmic reticulum stress in the in vitro ICH cell model. (A) Confocal images and three-dimensional reconstruction of endoplasmic reticulum continuity of neurons by ER tracker after hemin and rtPA treatment. Scale bars: 3 µm. (B–F) Quantitative analysis of ER stress–associated proteins of neurons. Data are shown as mean ± SEM ( n = 3 per group). * P < 0.05, ** P < 0.01, vs. control group; # P < 0.05, ## P < 0.01, vs . hemin group (one-way analysis of variance followed by Tukey’s post hoc test). (G) Immunofluorescence staining images of p-PERK (red, labeled by Cy3) in neurons after hemin and rtPA treatment. Scale bars: 50 µm. 3D: Three-dimensional; ATF6: cyclic AMP-dependent transcription factor ATF-6 alpha; DAPI: 4′,6-diamidino-2-phenylindole; eIF2α: eukaryotic translation initiation factor 2 subunit alpha; ER: endoplasmic reticulum; Grp78: endoplasmic reticulum chaperone BiP; PERK: PRKR-like endoplasmic reticulum kinase; rtPA: recombinant tissue plasminogen activator.

Techniques: In Vitro, Control, Immunofluorescence, Staining, Labeling, Recombinant

The PI3K inhibitor LY294002 reverses the anti-ER stress effect of rtPA and the EGF domain of rtPA may mediate the PI3K/AKT pathway in the ICH in vitro cell model. (A–C) Analysis of ER stress–associated proteins ( n = 3 per group). (D) Confocal images and three-dimensional reconstruction of endoplasmic reticulum continuity of neurons by ER tracker after rtPA and PI3K inhibitor LY294002 treatment. Scale bars: 3 µm. (E) Immunofluorescence images of p-PERK (red, labeled by Cy3) in neurons after rtPA and PI3K inhibitor LY294002 treatment. Scale bars: 50 µm. (F–H) Analysis of PI3K p85 and p-AKT. Data are represented as mean ± SEM ( n = 3 per group). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . hemin group; & P < 0.05, && P < 0.01, vs . hemin + rtPA group; # P < 0.05, vs . hemin + rtPA + DMSO group (one-way analysis of variance followed by Tukey’s post hoc test). (I) Transmission electron microscopy images of cells after rtPA and rtPA domain inhibitor treatment. Scale bar: 100 µm. 3D: Three-dimensional; AKT: RAC-alpha serine/threonine-protein kinase; ATF6: cyclic AMP-dependent transcription factor ATF-6 alpha; DAPI: 4′,6-diamidino-2-phenylindole; EGF: epidermal growth factor; eIF2α: eukaryotic translation initiation factor 2 subunit alpha; ER: endoplasmic reticulum; LY294002: PI3K inhibitor; mTOR: mammalian target of rapamycin; PERK: PRKR-like endoplasmic reticulum kinase; PI3K: phosphatidylinositol 3-kinase regulatory subunit alpha; 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: The PI3K inhibitor LY294002 reverses the anti-ER stress effect of rtPA and the EGF domain of rtPA may mediate the PI3K/AKT pathway in the ICH in vitro cell model. (A–C) Analysis of ER stress–associated proteins ( n = 3 per group). (D) Confocal images and three-dimensional reconstruction of endoplasmic reticulum continuity of neurons by ER tracker after rtPA and PI3K inhibitor LY294002 treatment. Scale bars: 3 µm. (E) Immunofluorescence images of p-PERK (red, labeled by Cy3) in neurons after rtPA and PI3K inhibitor LY294002 treatment. Scale bars: 50 µm. (F–H) Analysis of PI3K p85 and p-AKT. Data are represented as mean ± SEM ( n = 3 per group). * P < 0.05, ** P < 0.01, *** P < 0.001, vs . hemin group; & P < 0.05, && P < 0.01, vs . hemin + rtPA group; # P < 0.05, vs . hemin + rtPA + DMSO group (one-way analysis of variance followed by Tukey’s post hoc test). (I) Transmission electron microscopy images of cells after rtPA and rtPA domain inhibitor treatment. Scale bar: 100 µm. 3D: Three-dimensional; AKT: RAC-alpha serine/threonine-protein kinase; ATF6: cyclic AMP-dependent transcription factor ATF-6 alpha; DAPI: 4′,6-diamidino-2-phenylindole; EGF: epidermal growth factor; eIF2α: eukaryotic translation initiation factor 2 subunit alpha; ER: endoplasmic reticulum; LY294002: PI3K inhibitor; mTOR: mammalian target of rapamycin; PERK: PRKR-like endoplasmic reticulum kinase; PI3K: phosphatidylinositol 3-kinase regulatory subunit alpha; rtPA: recombinant tissue plasminogen activator.

Techniques: In Vitro, Immunofluorescence, Labeling, Transmission Assay, Electron Microscopy, Recombinant

TLR4/MyD88/PKCδ/SHP-1 signaling cascade induced podocyte damage through regulating ER stress in DKD and podocyte damage. (A) The protein levels of ER stress markers BIP, sXBP-1, ATF4 and ATF6 were determined in renal tissues from mice at 12 and 16 weeks by western blot. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01 ( n = 8). (B) Representative double IF staining of the expression ATF4 and podocyte marker synaptopodin in mice at 16 weeks. Scale bar, 20 μm ( n = 8). (C) Representative western blot to assess the levels of ER stress markers BIP, sXBP-1, ATF4 and ATF6 in cultured mouse podocytes from treatments of LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ### P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (D) ER morphological alterations and ER stress illustrated by ER-tracker staining in podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, or MCP-1 shRNAs, or pretreatment of 4-PBA. Scale bar, 10 μm. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; # P < 0.05 ( n = 3 independent experiments). (E) Representative western blot to detect the expression of ER stress proteins BIP, sXBP-1, ATF4 and ATF6 in cultured podocytes after HG stimulation for 48 h with or without transfection of PKCδ, SHP-1 or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA and data are presented as mean ± SD. ** P < 0.01; *** P < 0.01; **** P < 0.01; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments).

Journal: Journal of Advanced Research

Article Title: Podocyte TLR4 deletion alleviates diabetic kidney disease through prohibiting PKCδ/SHP-1-dependent ER stress and relieving podocyte damage and inflammation

doi: 10.1016/j.jare.2025.07.013

Figure Lengend Snippet: TLR4/MyD88/PKCδ/SHP-1 signaling cascade induced podocyte damage through regulating ER stress in DKD and podocyte damage. (A) The protein levels of ER stress markers BIP, sXBP-1, ATF4 and ATF6 were determined in renal tissues from mice at 12 and 16 weeks by western blot. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01 ( n = 8). (B) Representative double IF staining of the expression ATF4 and podocyte marker synaptopodin in mice at 16 weeks. Scale bar, 20 μm ( n = 8). (C) Representative western blot to assess the levels of ER stress markers BIP, sXBP-1, ATF4 and ATF6 in cultured mouse podocytes from treatments of LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ### P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (D) ER morphological alterations and ER stress illustrated by ER-tracker staining in podocytes treated with LG or HG for 48 h with or without transfection of TLR4, PKCδ, or MCP-1 shRNAs, or pretreatment of 4-PBA. Scale bar, 10 μm. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ## P < 0.01; # P < 0.05 ( n = 3 independent experiments). (E) Representative western blot to detect the expression of ER stress proteins BIP, sXBP-1, ATF4 and ATF6 in cultured podocytes after HG stimulation for 48 h with or without transfection of PKCδ, SHP-1 or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA and data are presented as mean ± SD. ** P < 0.01; *** P < 0.01; **** P < 0.01; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments).

Article Snippet: The following primary antibodies were used: f4/80 (29414–1-AP), desmin (16520–1-AP), MCP-1 (26161–1-AP), podocin (20384–1-AP), synaptopodin (21064–1-AP), TLR4 (66350–1-Ig), β-actin (66009–1-Ig), PKCδ(14188–1-AP), ATF6 (24169–1-AP), p-cadherin (13773–1-AP) were from Proteintech (IL, USA); phospho-PKCδ (Y311) (ab76181), PKCdelta (ab182126), SHP-1 (ab227503), HA (ab9110) were from Abcam (MA, USA); BIP (M010300), MyD88 (P012343) were from Epizyme, Shanghai, China); and ATF4 (11815), sXBP-1 (40435) were from Cell Signaling Technology (MA, USA).

Techniques: Western Blot, Staining, Expressing, Marker, Cell Culture, Transfection

Further validation of PKCδ activation being indispensable for ER stress and MCP-1 production through SHP-1 in diabetic podocyte injury. (A) Schematic representation of PKCδ plasmids to express PKCδ proteins in DN (tyrosine mutation at amino acid 311), CA or WT form. (B) Representative western blot to validate the expression of DN-PKCδ, CA-PKCδ and WT-PKCδ plasmids in HEK-293 T cells by using antibodies specific for HA-tag, with GAPDH detection as a loading control. ( n = 3 independent experiments). (CDE) Determination of protein levels of SHP-1 (C), ER stress proteins BIP, sXBP-1, ATF4 and ATF6 (D), and MCP-1 (E) in cultured mouse podocytes transfected with DN-, CA- and WT-PKCδ plasmids, or control plasmids under LG or HG conditions. P values were determined by one-way ANOVA and data are presented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.01; **** P < 0.01; ## P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments).

Journal: Journal of Advanced Research

Article Title: Podocyte TLR4 deletion alleviates diabetic kidney disease through prohibiting PKCδ/SHP-1-dependent ER stress and relieving podocyte damage and inflammation

doi: 10.1016/j.jare.2025.07.013

Figure Lengend Snippet: Further validation of PKCδ activation being indispensable for ER stress and MCP-1 production through SHP-1 in diabetic podocyte injury. (A) Schematic representation of PKCδ plasmids to express PKCδ proteins in DN (tyrosine mutation at amino acid 311), CA or WT form. (B) Representative western blot to validate the expression of DN-PKCδ, CA-PKCδ and WT-PKCδ plasmids in HEK-293 T cells by using antibodies specific for HA-tag, with GAPDH detection as a loading control. ( n = 3 independent experiments). (CDE) Determination of protein levels of SHP-1 (C), ER stress proteins BIP, sXBP-1, ATF4 and ATF6 (D), and MCP-1 (E) in cultured mouse podocytes transfected with DN-, CA- and WT-PKCδ plasmids, or control plasmids under LG or HG conditions. P values were determined by one-way ANOVA and data are presented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.01; **** P < 0.01; ## P < 0.01; # P < 0.05; ## P < 0.01; ### P < 0.01; #### P < 0.01 ( n = 3 independent experiments).

Techniques: Biomarker Discovery, Activation Assay, Mutagenesis, Western Blot, Expressing, Control, Cell Culture, Transfection

TLR4/MyD88/PKCδ/SHP-1 signaling-dependent ER stress stimulated MCP-1 production via enhanced transcription activity by ATF4 binding to its promoter in hyperglycemic podocytes. (A) The altered expression of MCP-1 in renal tissue from mice at 12 and 16 weeks by western blot. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01 ( n = 8 samples). (B) Representative western blot to assess the levels of MCP-1 in cultured mouse podocytes from treatments of LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (C) Representative western blot to show the expression of MCP-1 in podocytes after HG stimulation for 48 h with or without transfection of PKCδ, SHP-1 or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA and data are presented as mean ± SD. ** P < 0.01; *** P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (DE) The expression of MCP-1 by western blot (D), and ER stress markers BIP, sXBP-1, ATF4 and ATF6 (E) after HG stimulation for 48 h with or without MCP-1 or scrambled shRNA transfection. P values were determined by one-way ANOVA and data are presented as mean ± SD. ## P < 0.01; #### P < 0.01 ( n = 3 independent experiments). (F) Schematic description of primers spanning the distal, medial, and proximal regions of the MCP-1 promoter, encompassing predicted binding sites in the present study. (GH) ChIP analysis using anti-ATF4 or anti-IgG antibodies were performed either in cultured mouse podocytes after LG or HG treatment for 48 h (G), or HG stimulation with transfection of TLR4, PKCδ, MCP-1 or scrambled shRNA, or pretreatment of 4-PBA (H). Real-time PCR was used for the detection of the ChIP signals. P values were determined by Student’s t -test, or by one-way ANOVA, and data are presented as mean ± SD. * P < 0.05; ** P < 0.01; **** P < 0.01; #### P < 0.01 ( n = 3 independent experiments). (I) Determination of ATF4 nuclear accumulation in podocytes under LG or HG conditions by western blot. P values were determined by Student’s t -test and data are presented as mean ± SD. ** P < 0.01; ## P < 0.01 ( n = 3 independent experiments). (J) Assessment of nuclear ATF4 protein levels in cultured podocytes transfected with TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreated with 4-PBA under HG conditions for 48 h. P values were determined by one-way ANOVA and data are presented as mean ± SD. * P < 0.05; * P < 0.01 ( n = 3 independent experiments).

Journal: Journal of Advanced Research

Article Title: Podocyte TLR4 deletion alleviates diabetic kidney disease through prohibiting PKCδ/SHP-1-dependent ER stress and relieving podocyte damage and inflammation

doi: 10.1016/j.jare.2025.07.013

Figure Lengend Snippet: TLR4/MyD88/PKCδ/SHP-1 signaling-dependent ER stress stimulated MCP-1 production via enhanced transcription activity by ATF4 binding to its promoter in hyperglycemic podocytes. (A) The altered expression of MCP-1 in renal tissue from mice at 12 and 16 weeks by western blot. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; # P < 0.05; ## P < 0.01 ( n = 8 samples). (B) Representative western blot to assess the levels of MCP-1 in cultured mouse podocytes from treatments of LG or HG for 48 h with or without transfection of TLR4, MyD88 or scrambled shRNAs. P values were determined by one-way ANOVA and data are presented as mean ± SD. **** P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (C) Representative western blot to show the expression of MCP-1 in podocytes after HG stimulation for 48 h with or without transfection of PKCδ, SHP-1 or scrambled shRNAs, or pretreatment of 4-PBA. P values were determined by one-way ANOVA and data are presented as mean ± SD. ** P < 0.01; *** P < 0.01; ### P < 0.01 ( n = 3 independent experiments). (DE) The expression of MCP-1 by western blot (D), and ER stress markers BIP, sXBP-1, ATF4 and ATF6 (E) after HG stimulation for 48 h with or without MCP-1 or scrambled shRNA transfection. P values were determined by one-way ANOVA and data are presented as mean ± SD. ## P < 0.01; #### P < 0.01 ( n = 3 independent experiments). (F) Schematic description of primers spanning the distal, medial, and proximal regions of the MCP-1 promoter, encompassing predicted binding sites in the present study. (GH) ChIP analysis using anti-ATF4 or anti-IgG antibodies were performed either in cultured mouse podocytes after LG or HG treatment for 48 h (G), or HG stimulation with transfection of TLR4, PKCδ, MCP-1 or scrambled shRNA, or pretreatment of 4-PBA (H). Real-time PCR was used for the detection of the ChIP signals. P values were determined by Student’s t -test, or by one-way ANOVA, and data are presented as mean ± SD. * P < 0.05; ** P < 0.01; **** P < 0.01; #### P < 0.01 ( n = 3 independent experiments). (I) Determination of ATF4 nuclear accumulation in podocytes under LG or HG conditions by western blot. P values were determined by Student’s t -test and data are presented as mean ± SD. ** P < 0.01; ## P < 0.01 ( n = 3 independent experiments). (J) Assessment of nuclear ATF4 protein levels in cultured podocytes transfected with TLR4, PKCδ, MCP-1 or scrambled shRNAs, or pretreated with 4-PBA under HG conditions for 48 h. P values were determined by one-way ANOVA and data are presented as mean ± SD. * P < 0.05; * P < 0.01 ( n = 3 independent experiments).

Techniques: Activity Assay, Binding Assay, Expressing, Western Blot, Cell Culture, Transfection, shRNA, Real-time Polymerase Chain Reaction

Review

Structured Review

Images

1) Product Images from "PRAS40 activates the IRE1α-XBP-1-mediated unfolded protein response to exacerbate colorectal cancer by enhancing ST6Gal1-dependent α-2, 6 sialylation of GRP78"

Similar Products

Image Search Results