Journal: The Journal of Biological Chemistry

Article Title: Tubulin hyperacetylation drives HMGB1 nuclear exit via the ROS-PARP1 axis, leading to rotenone-induced G2/M arrest

doi: 10.1016/j.jbc.2025.110695

Figure Lengend Snippet: Rotenone triggers time-dependent HMGB1 nuclear exit coupled with increased PARylation. A , representative confocal images of time-dependent HMGB1 ( green ) localization in SH-SY5Y cells treated with rotenone (5μM) or DMSO control. DAPI ( blue ) indicates nuclear staining (scale bar = 10 μm). B , quantification of SH-SY5Y cells with cytoplasmic enrichment of HMGB1 shown in ( A ). Nucleus was defined by area of DAPI. Cytosolic HMGB1 was calculated by subtracting nuclear region of interest (ROI) from the area of green channel. n = 100 cells for each quantification. C , immunoblot analysis of HMGB1 protein levels in the nuclear and cytosolic fraction of SH-SY5Y cells treated with rotenone for 2, 4, 6 and 24 h and compared with DMSO treated controls. GAPDH was used as cytosolic loading control and lamin A/C was used as a nuclear loading control ( left panel ). The nuclear to cytoplasmic ratio was quantified using the lower band obtained in the cytoplasmic fraction of the gels ( right panel ). D , co-immunoprecipitation analysis was performed to assess the PARylation status of HMGB1 in whole-cell extracts following 24 h of rotenone treatment. Cells were immunoprecipitated with an anti-HMGB1 antibody, and PAR ( upper panel ) and HMGB1 ( middle panel ; low and high exposures) levels were analyzed. Additionally, cells were immunoprecipitated with an anti-PAR antibody, and HMGB1 levels ( lower panel ) were examined. E , immunoblot analysis of HMGB1 protein levels in the nuclear and cytosolic fraction of cells treated with rotenone for 24 h in the presence or absence of the PARP inhibitor PJ34 (50μM). GAPDH was used as cytosolic loading control and lamin A/C was used as a nuclear loading control ( left panel ). The nuclear to cytoplasmic ratio was quantified using the lower band obtained in the cytoplasmic fraction of the gels ( right panel ). F , representative confocal images of HMGB1 ( green ) and DAPI (blue) in SH-SY5Y cells treated with rotenone or DMSO control in the presence of PJ34. DAPI ( blue ) indicates nuclear staining (scale bar = 10 μm). G , quantification of cells with cytoplasmic enrichment of HMGB1. n = 100 cells for each quantification. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparison test (∗∗∗ p < 0.001; n = 3 biological replicates).

Article Snippet: For determining the level of PARylation, anti-PAR antibody (#MCA-1480) and mouse monoclonal antibody for β-actin (#MCA5775GA) was obtained from Bio-Rad.

Techniques: Control, Staining, Western Blot, Immunoprecipitation, Comparison

Journal: Cell reports

Article Title: Deciphering the functional landscape of phosphosites with deep neural network.

doi: 10.1016/j.celrep.2023.113048

Figure Lengend Snippet: Figure 4. The predicted functional score and entropy value for phosphosites in NADK and the in vitro enzymatic acitivity for NADK-WT and NADK mutants (A) Functional score for phosphosites identified in NADK with unknown function (blue), known activity (brown), and predicted activity (yellow). (B) The entropy value for residues in NADK, with phosphosites colored in blue and yellow. (C) Four functional phosphorylated serine sites in NADK are conserved across different species. (D) NADK-WT (wild type), NADK-S48/50D (S48/50 were both mutated to aspartic acids), NADK-S44/ 46D (S44/46 were both mutated to aspartic acids), and NADK-S44/46/48/50D (S44/46/48/50 were mutated to aspartic acids) proteins were expressed and purified in BL21-E. coli bacteria. In vitro enzy- matic activity was tested, and the graph was per- formed using substrate (NAD+) concentration and the reaction initial rate V0.

Article Snippet: For in vitro kinase assay followed by NADK enzymatic assay, 50mg His-NADK protein was incubated with 20 mg GST-ERK2 or ERK2-K54R and 10mg constitutively active GST-MEK1 (S218/222D mutation with deletion of aa-No.32-51) in kinase assay buffer with or without ERK inhibitor GDC-0994(HY-15947, MedChemExpress) for 1-2h at room temperature.

Techniques: Functional Assay, In Vitro, Activity Assay, Bacteria, Concentration Assay

Journal: Cell reports

Article Title: Deciphering the functional landscape of phosphosites with deep neural network.

doi: 10.1016/j.celrep.2023.113048

Figure Lengend Snippet: Figure 5. ERK1/2 kinases interact with NADK (A and B) FLAG-tagged ERK1 or ERK2 and myc-tagged NADK plasmids were transfected into 293T cells, and coIP assay was performed using FLAG-M2 beads. (C) FLAG-tagged ERK2-, JNK1-, and P38a-expressing plasmids were co-transfected into 293T cells with myc-tagged NADK plasmids, and coIP assay was performed using FLAG-M2 beads. (D) FLAG-tagged NADK plasmids were transfected into 293T cells, coIP assay was performed using FLAG-M2 beads, and endogenous ERK1/2 was detected using ERK1/2 antibody. (E and F) His-tagged NADK and GST-tagged ERK2 were expressed and purified using BL21-E. coli bacteria, and His pull-down and GST pull-down were performed using His beads and GSH beads. (G) Microscale thermophoresis assay (MST) was performed using purified His-NADK (200 nM) and gradient-diluted ERK2 protein. Three independent MST assays were performed to identify Kd, which is shown as mean ± SD.

Article Snippet: For in vitro kinase assay followed by NADK enzymatic assay, 50mg His-NADK protein was incubated with 20 mg GST-ERK2 or ERK2-K54R and 10mg constitutively active GST-MEK1 (S218/222D mutation with deletion of aa-No.32-51) in kinase assay buffer with or without ERK inhibitor GDC-0994(HY-15947, MedChemExpress) for 1-2h at room temperature.

Techniques: Transfection, Co-Immunoprecipitation Assay, Expressing, Bacteria, Microscale Thermophoresis

Journal: Cell reports

Article Title: Deciphering the functional landscape of phosphosites with deep neural network.

doi: 10.1016/j.celrep.2023.113048

Figure Lengend Snippet: Figure 6. ERK2 phosphorylates NADK-S48/50 and activates NADK enzymatic activity (A) Constitutively active GST-MEK1 (S218/222D mutation with deletion of aa 32–51) and GST-ERK2 or GST-ERK2-kinase-dead protein and His-NADK protein were incubated with ATP-g-S and ERK inhibitor GDC-0994 or DMSO for 1 to 2 h. PNBM (final concentration was 2.5 mM) was added to alkylate the proteins, and then His-NADK was isolated using Ni beads, and western blotting was performed to detect NADK phosphorylation using thiophosphate ester specific antibody. (B) Constitutively active GST-MEK1 (S218/222D mutation with deletion of aa 32–51) and GST-ERK2 were incubated with mutant His-NADK (mutation from serine to alanine) for in vitro kinase assay using ATP-g-S, and western blotting was performed to detect NADK phosphorylation. (C) His-NADK was incubated with constitutively active GST-MEK1 and GST-ERK2 with or without 0.5 mM ERK inhibitor GDC-0994 for in vitro kinase assay. Phosphorylated His-NADK was isolated by using Ni beads, and an in vitro NADK enzymatic assay was carried out. The graph was performed using substrate (NAD+) concentration and the reaction initial rate V0. (D) His-NADK was incubated with constitutively active GST-MEK1 and GST-ERK2 or kinase-dead-mutant GST-ERK2(K54R) for in vitro kinase assay, and then NADK was isolated, and in vitro enzymatic assay was carried out. The graph was performed using substrate (NAD+) concentration and initial rate V0. (E) His-NADK-S48/50A mutant protein was incubated with constitutively active GST-MEK1 and GST-ERK2 for in vitro kinase assay, and then NADK was isolated, and in vitro enzymatic assay was carried out. The graph was performed using substrate (NAD+) concentration and the reaction initial rate V0. Error bars denote the mean ± SD of experiments completed in three replicates. (F) ERK1/2 kinase phosphorylates NADK-S48/50 and activates NADK activity to enhance NADP+/NADPH synthesis.

Article Snippet: For in vitro kinase assay followed by NADK enzymatic assay, 50mg His-NADK protein was incubated with 20 mg GST-ERK2 or ERK2-K54R and 10mg constitutively active GST-MEK1 (S218/222D mutation with deletion of aa-No.32-51) in kinase assay buffer with or without ERK inhibitor GDC-0994(HY-15947, MedChemExpress) for 1-2h at room temperature.

Techniques: Activity Assay, Mutagenesis, Incubation, Concentration Assay, Isolation, Western Blot, Phospho-proteomics, In Vitro, Kinase Assay, Enzymatic Assay

Journal: Cancer Management and Research

Article Title: PUF60 accelerates the progression of breast cancer through downregulation of PTEN expression

doi: 10.2147/CMAR.S180242

Figure Lengend Snippet: PUF60 was overexpressed in breast cancer. Notes: ( A ) Oncomine database was used to analyze PUF60 expression in patients with breast cancer or healthy individuals. ( B, C ) RT-PCR together with Western blotting technologies was recruited to determine the mRNA and protein expression of PUF60 in 30 matched breast cancer tissues and normal tissues ( ** P <0.01). ( D ) The expression of PUF60 in 15 paired breast cancer tissues and normal tissues was detected by immunohistochemistry, and the score of PUF60 was recorded. ( E, F ) RT-PCR and Western blotting technologies were recruited to assess the mRNA and protein expression of PUF60 in breast normal MCF-10A cells and breast cancer MDA-MB-231, HCC1937, and MCF-7 cells. These experiments were repeated three times ( *** P <0.001). ( G ) RT-PCR was used to evaluate the mRNA expression of PUF60 in the blood samples from 30 cases of breast cancer patients or 30 cases of healthy individuals. Abbreviations: IHC, immunohistochemstry; RT-PCR, real-time PCR.

Article Snippet: To upregulate or downregulate the expression of PUF60, lentivirus containing PUF60 open reading frame (OE-PUF60) and shRNA targeting human PUF60 gene were purchased from OriGene (Beijing, China).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Real-time Polymerase Chain Reaction

Journal: Cancer Management and Research

Article Title: PUF60 accelerates the progression of breast cancer through downregulation of PTEN expression

doi: 10.2147/CMAR.S180242

Figure Lengend Snippet: Relationship of PUF60 expression and breast cancer in patients’ clinical and pathological characteristics

Article Snippet: To upregulate or downregulate the expression of PUF60, lentivirus containing PUF60 open reading frame (OE-PUF60) and shRNA targeting human PUF60 gene were purchased from OriGene (Beijing, China).

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: PUF60 accelerates the progression of breast cancer through downregulation of PTEN expression

doi: 10.2147/CMAR.S180242

Figure Lengend Snippet: Relationship of PUF60 expression and breast cancer patients’ overall survival. Note: Kaplan–Meier analysis was recruited to evaluate the overall survival of breast cancer patients with PUF60 high expression (n=48) and low expression (n=30).

Article Snippet: To upregulate or downregulate the expression of PUF60, lentivirus containing PUF60 open reading frame (OE-PUF60) and shRNA targeting human PUF60 gene were purchased from OriGene (Beijing, China).

Techniques: Expressing

Journal: Cancer Management and Research

Article Title: PUF60 accelerates the progression of breast cancer through downregulation of PTEN expression

doi: 10.2147/CMAR.S180242

Figure Lengend Snippet: Upregulation of PUF60 enhanced the growth and inhibited the apoptosis of breast cancer cells. Notes: MDA-MB-231 and HCC1937 cells were infected with sh-PUF60, sh-NC, OE-PUF60, or OE-NC, then ( A, B ) MDA-MB-231 and HCC1937 cells were collected and submitted to RT-PCR and Western blotting experiments to detect the overexpressing or knockdown efficiency of OE-PUF60 or sh-PUF60 after 48 hours of the infection. ( C, D ) CCK-8 assay was performed to determine cell growth after 1, 2, 3, 4, or 5 days of the above treatments. ( E ) Annexin V/PI apoptosis detection kit was used to analyze the apoptosis of MDA-MB-231 or HCC1937 cells after 48 hours of cell infection. ( F, G ) The expression levels of Bcl-2 and Bax proteins were detected by Western blotting analysis in MDA-MB-231 and HCC1937 cells. All experiments were repeated three times ( ** P <0.01, *** P <0.001, sh-PUF60 group compared to sh-NC group; # P <0.05, ## P <0.01, ### P <0.001, OE-PUF60 group compared to OE-NC group). Abbreviations: PI, propidium iodide; RT-PCR, real-time PCR.

Article Snippet: To upregulate or downregulate the expression of PUF60, lentivirus containing PUF60 open reading frame (OE-PUF60) and shRNA targeting human PUF60 gene were purchased from OriGene (Beijing, China).

Techniques: Infection, Reverse Transcription Polymerase Chain Reaction, Western Blot, Knockdown, CCK-8 Assay, Expressing, Real-time Polymerase Chain Reaction

Journal: Cancer Management and Research

Article Title: PUF60 accelerates the progression of breast cancer through downregulation of PTEN expression

doi: 10.2147/CMAR.S180242

Figure Lengend Snippet: Upregulation of PUF60 significantly improved the migration and invasion of breast cancer cells. Notes: MDA-MB-231 and HCC1937 cells were infected with sh-PUF60, sh-NC, OE-PUF60, or OE-NC, then ( A ) Transwell chambers with Matrigel coated were recruited to determine the invasion of MDA-MB-231 and HCC1937 cells after 48 hours of cell infection. ( B ) Transwell chambers without Matrigel coated were used to detect the migration abilities of MDA-MB-231 and HCC1937 cells after 24 hours of cell infection. All experiments were repeated three times ( ** P <0.01, *** P <0.001, sh-PUF60 group compared to sh-NC group; # P <0.05, ## P <0.01, ### P <0.001, OE-PUF60 group compared to OE-NC group).

Article Snippet: To upregulate or downregulate the expression of PUF60, lentivirus containing PUF60 open reading frame (OE-PUF60) and shRNA targeting human PUF60 gene were purchased from OriGene (Beijing, China).

Techniques: Migration, Infection

Journal: Cancer Management and Research

Article Title: PUF60 accelerates the progression of breast cancer through downregulation of PTEN expression

doi: 10.2147/CMAR.S180242

Figure Lengend Snippet: Upregulation of PUF60 activated the AKT/mTOR/PTEN signaling transduction pathway in breast cancer cells. Notes: MDA-MB-231 and HCC1937 cells were infected with OE-PUF60 or OE-NC for 48 hours, then cells were harvested and submitted to ( A, B ) Western blotting to detect the expression of AKT, p-AKT, mTOR, PI3K, and PTEN. ( C, D ) RT-PCR to assess the mRNA levels of AKT, mTOR, PI3K, and PTEN. ( E ) IP assay was used to test the interaction between PUF60 protein and AKT, mTOR, or PTEN protein. All experiments were repeated three times ( ** P <0.01, *** P <0.001). Abbreviation: IP, immunoprecipitation.

Article Snippet: To upregulate or downregulate the expression of PUF60, lentivirus containing PUF60 open reading frame (OE-PUF60) and shRNA targeting human PUF60 gene were purchased from OriGene (Beijing, China).

Techniques: Transduction, Infection, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction, Immunoprecipitation

Journal: Cancer Management and Research

Article Title: PUF60 accelerates the progression of breast cancer through downregulation of PTEN expression

doi: 10.2147/CMAR.S180242

Figure Lengend Snippet: Upregulation of PUF60 weakened the stability of PTEN and facilitated its ubiquitination. Notes: ( A ) Cell immunofluorescence technique was carried out to determine the location of PUF60 and PTEN protein in MDA-MB-231 and HCC1937 cells. ( B ) MDA-231 and HCC1937 cells were given sh-NC, sh-PUF60, OE-NC, or OE-PUF60 for 24 hours, then 100 ng/mL cyclohexamide (CHX) was added to the culture medium to stop protein synthesis, then cells were collected and submitted to Western blotting assay with antibody against PTEN as the indicated times. ( C ) IP assay was used to assess the effects of PUF60 upregulation in the ubiquitination of PTEN. All experiments were repeated three times. Abbreviation: IP, immunoprecipitation.

Article Snippet: To upregulate or downregulate the expression of PUF60, lentivirus containing PUF60 open reading frame (OE-PUF60) and shRNA targeting human PUF60 gene were purchased from OriGene (Beijing, China).

Techniques: Ubiquitin Proteomics, Immunofluorescence, Western Blot, Immunoprecipitation

Journal: Cancer Management and Research

Article Title: PUF60 accelerates the progression of breast cancer through downregulation of PTEN expression

doi: 10.2147/CMAR.S180242

Figure Lengend Snippet: Upregulation promoted the tumorigenesis of breast cancer cells. Note: In vivo tumor formation assay was carried out to detect the effects of PUF60/PTEN on the tumorigenesis of MDA-MB-231 and HCC1937 cells, with six nude mice in each group.

Article Snippet: To upregulate or downregulate the expression of PUF60, lentivirus containing PUF60 open reading frame (OE-PUF60) and shRNA targeting human PUF60 gene were purchased from OriGene (Beijing, China).

Techniques: In Vivo, Tube Formation Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Acteoside ameliorates hepatocyte ferroptosis and hepatic ischemia-reperfusion injury via targeting PCBP2

doi: 10.1016/j.apsb.2025.03.002

Figure Lengend Snippet: ACT upregulates the expression of PCBP2 in vivo and in vitro . (A) The heatmap of ferroptosis-related genes of mice liver. (B) The protein levels of PCBP2 were measured by Western blot and normalized by β -actin in mice liver. (C) Representative images of PCBP2 staining in the mouse liver, scale bar = 40 μm. (D) The volcano plot of ferroptosis-related genes in hepatocytes. (E) Relative mRNA levels of Pcbp2 in AML12 cells were measured by qPCR and further normalized with hprt1 . Statistical significance: # P < 0.05, ### P < 0.001, compared with relative model groups ( n = 6). (F) The protein levels of PCBP2 were measured by Western blot and normalized by β -actin in the AML 12 cells. (G, H) Representative images of PCBP2 and ROS staining in the AML12 cells under HR or erastin insult, scale bar = 100 μm.

Article Snippet: The recombinant PCBP2 protein (CSB-EP622995HU) was applied both in vivo and in vitro (CUSABIO, Wuhan, China).

Techniques: Expressing, In Vivo, In Vitro, Western Blot, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: Acteoside ameliorates hepatocyte ferroptosis and hepatic ischemia-reperfusion injury via targeting PCBP2

doi: 10.1016/j.apsb.2025.03.002

Figure Lengend Snippet: ACT improves the expression and function of the System Xc – under hypoxia conditions. (A) The circle chart (left panel, whole livers RNA-sequence data) and coefficient chart (middle and right panel, only hepatocytes RNA-sequence data) for the relationship between PCBP2 and SLC3A2/SLC7A11. (B) Relative mRNA levels of Slc3a2 , Slc7a11 and Gpx4 in mouse livers were measured by qPCR and further normalized with hprt1 . Statistical significance: ∗ P < 0.05, compared with relative control groups; ### P < 0.001, compared with the relative model groups ( n = 6). (C)The protein levels of SLC3A2 and SLC7A11 were measured by Western blot and normalized by β -actin in mouse livers. (D) Representative images of SLC3A2 and SLC7A11 staining in the mouse liver, scale bar = 40 μm. Statistical significance: ∗∗∗ P < 0.001, compared with relative control groups; ### P < 0.001, compared with the relative model groups ( n = 6). (E) The content of Cys and GSH in mice liver. Statistical significance: ∗∗∗ P < 0.001, compared with relative control groups; ### P < 0.001, compared with the relative model groups ( n = 6). (F) Relative mRNA levels of Slc3a2 and Slc7a11 in AML 12 cells were measured by qPCR and further normalized with hprt1 . Statistical significance: ∗∗ P < 0.01, ∗∗∗ P < 0.001, compared with relative control groups; # P < 0.05, ## P < 0.01, ### P < 0.001, compared with the relative model groups. (G) The protein levels of SLC3A2 and SLC7A11 were measured by Western blot and normalized by β -actin in AML 12 cells. (H) Representative images of SLC3A2 and SLC7A11 staining in the AML 12 cells, scale bar = 100 μm. (I) The quantization chart for SLC3A2 and SLC7A11 staining in AML 12 cells. (J) The content of Cys and GSH in AML 12 cells. Statistical significance: ∗∗∗ P < 0.001, compared with relative control groups; # P < 0.05, ## P < 0.01, ### P < 0.001, compared with the relative model groups.

Article Snippet: The recombinant PCBP2 protein (CSB-EP622995HU) was applied both in vivo and in vitro (CUSABIO, Wuhan, China).

Techniques: Expressing, Sequencing, Control, Western Blot, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: Acteoside ameliorates hepatocyte ferroptosis and hepatic ischemia-reperfusion injury via targeting PCBP2

doi: 10.1016/j.apsb.2025.03.002

Figure Lengend Snippet: ACT promotes PCBP2 to maintain the stability of system Xc – . (A) The SPR detection of ACT with PCBP2 protein. (B) The MST results of ACT with PCBP2 protein. (C) The molecular simulation docking between ACT and PCBP2–SLC3A2 protein complex. (D) The molecular simulation docking between ACT and PCBP2–SLC7A11 protein complex. (E) The co-IP results between PCBP2 and SLC3A2/SLC7A11. (F) The representative images of multi-color fluorescence staining of PCBP2/SLC3A2/SLC7A11 in mouse livers, scale bar = 40 μm. (G) The mRNA changes of Pcbp2 , Slc3a2 and Slc7a11 in hepatocytes. (H) Relative mRNA levels of Slc3a2 , Slc7a11 and PcBP2 in AML12 cells were measured by qPCR and further normalized with hprt1 . Statistical significance: ∗∗∗ P < 0.001, compared with control group; # P < 0.05, ## P < 0.01, ### P < 0.001, compared with the siPCBP2 group ( n = 3). (I) The protein levels of PCBP2, SLC3A2 and SLC7A11 were measured by Western blot and normalized by β -actin in AML 12 cells. (J) The content of Cys, MDA and SOD in AML 12 cells. Statistical significance: ∗∗∗ P < 0.001, compared with control group; # P < 0.05, ### P < 0.001, compared with the siPCBP2 group ( n = 3).

Article Snippet: The recombinant PCBP2 protein (CSB-EP622995HU) was applied both in vivo and in vitro (CUSABIO, Wuhan, China).

Techniques: Co-Immunoprecipitation Assay, Fluorescence, Staining, Control, Western Blot

Journal: Acta Pharmaceutica Sinica. B

Article Title: Acteoside ameliorates hepatocyte ferroptosis and hepatic ischemia-reperfusion injury via targeting PCBP2

doi: 10.1016/j.apsb.2025.03.002

Figure Lengend Snippet: ACT upregulates PCBP2 to inhibit the P300/HIF1 α /HMGB1 signaling in hepatocytes. (A) The diagram of pathway enrichment analysis. (B) The ridgeline plot (left) of DAMPs related genes and the coefficient diagram (right) of HMGB1 and PCBP2. (C) The level of HMGB1 in mouse serum and AML 12 cell medium. Statistical significance: ∗∗∗ P < 0.001, compared with the control group; ## P < 0.01, ### P < 0.001, compared with the relative model groups ( n = 6). (D) The RBP-mRNA CLIP results of PCBP2–HIF1 α , PCBP2–P300 and PCBP2–P300. (E) The mRNA changes of Hif1a , Ep300 and Hmgb1 in AML12 cells were measured by qPCR and further normalized with hprt1 . Statistical significance: ∗∗∗ P < 0.001, compared with the control group; # P < 0.05, ## P < 0.01, ### P < 0.001, compared with the relative model groups ( n = 3). (F) The protein levels of P300 and HMGB1 were measured by Western blot and normalized by β -actin in AML 12 cells. (G) The representative images of P300 and HMGB1 staining in AML 12 cells, scale bar = 100 μm.

Article Snippet: The recombinant PCBP2 protein (CSB-EP622995HU) was applied both in vivo and in vitro (CUSABIO, Wuhan, China).

Techniques: Control, Western Blot, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: Acteoside ameliorates hepatocyte ferroptosis and hepatic ischemia-reperfusion injury via targeting PCBP2

doi: 10.1016/j.apsb.2025.03.002

Figure Lengend Snippet: The hepatocyte specific knockout of PCBP2 interferes the therapeutic effects of ACT. (A) The particle size, (B) PDI, zeta potential and EE (%) of LNPs-si-PCBP2. (C) The stability of LNPs-si-PCBP2 in PBS and cell culture media. (D) The release behaviors of free si-RNA and LNPs-si-PCBP2. (E) Flow chart of ApoE-LNPs animal experiment. (F) The content of Fe 3+ , Fe 2+ , Cys and GSH in mouse livers. Statistical significance: ∗∗ P < 0.01, ∗∗∗ P < 0.001, compared with the sham group ( n = 6). Representative images of (G) H & E staining (scale bar = 40 μm), (H) PPB staining (scale bar = 40 μm), (I) immunofluorescence staining for PCBP2/SLC3A2/SLC7A11 (scale bar = 40 μm), (J) immunofluorescence staining for HMGB1 (scale bar = 40 μm) and (K) CD68 (scale bar = 40 μm) in mouse livers. (L) The Relative mRNA levels of slc3a2 , slc7a11 , p300 , hmgb1 and F4/80 were measured by qPCR and further normalized with hprt1 in mouse livers. Statistical significance: ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, compared with the sham group ( n = 6).

Article Snippet: The recombinant PCBP2 protein (CSB-EP622995HU) was applied both in vivo and in vitro (CUSABIO, Wuhan, China).

Techniques: Knock-Out, Zeta Potential Analyzer, Cell Culture, Staining, Immunofluorescence

Journal: International Journal of Molecular Sciences

Article Title: Neuroprotective Properties of Picroside II in a Rat Model of Focal Cerebral Ischemia

doi: 10.3390/ijms11114580

Figure Lengend Snippet: The number of PARP positive cells ( x̄ ± s).

Article Snippet: Rabbit anti-rat Caspase-3 and PARP moloclonal antibody, SABC immunohistochemistry kit, DAB dye were purchased from Boster Biological Company, Wuhan, China.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Neuroprotective Properties of Picroside II in a Rat Model of Focal Cerebral Ischemia

doi: 10.3390/ijms11114580

Figure Lengend Snippet: PARP positive cells in cortex shown by immunohistochemical assay × 400. The expression of PARP was very weak in the sham-operation group ( A ), increased in control group ( B ) and decreased in treatment group ( C ) significantly. Bar 25 μm.

Article Snippet: Rabbit anti-rat Caspase-3 and PARP moloclonal antibody, SABC immunohistochemistry kit, DAB dye were purchased from Boster Biological Company, Wuhan, China.

Techniques: Immunohistochemical staining, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Neuroprotective Properties of Picroside II in a Rat Model of Focal Cerebral Ischemia

doi: 10.3390/ijms11114580

Figure Lengend Snippet: The concentration in brain tissue of caspase-3 and PARP ( x̄ ± s).

Article Snippet: Rabbit anti-rat Caspase-3 and PARP moloclonal antibody, SABC immunohistochemistry kit, DAB dye were purchased from Boster Biological Company, Wuhan, China.

Techniques: Concentration Assay, Control