Journal: Frontiers in Immunology

Article Title: Foot-and-Mouth Disease Virus Counteracts on Internal Ribosome Entry Site Suppression by G3BP1 and Inhibits G3BP1-Mediated Stress Granule Assembly via Post-Translational Mechanisms

doi: 10.3389/fimmu.2018.01142

Figure Lengend Snippet: Confirmation of differentially expressed proteins and phosphoproteins by western blotting and Phos-tag western Blotting. (A) Analysis of ubiquitin conjugating enzyme E2 I, ubiquitin conjugating enzyme E2 L3, glyceraldehyde-3-phosphate dehydrogenase, β-actin expression levels in foot-and-mouth disease virus (FMDV)-infected and control cells by western blotting. SILAC-ratios and immunoblotting ratios (infection/control) were shown on the right side. (B) Analysis of the dynamic phosphorylation alterations of the three differentially phosphoproteins (ribosomal protein L15, chromosome 5 open reading frame 24, and FOS-like 2) in FMDV-infected and control cells by Phos-tag western blotting.

Article Snippet: To confirm the expression levels of GAPDH, ubiquitin conjugating enzyme E2 I (UBE2I), ubiquitin conjugating enzyme E2 L3 (UBE2L3), ribosomal protein L15 (RPL15), chromosome 5 open reading frame 24 (C5ORF24) and FOS-like 2 (FOSL2), anti-GAPDH antibody (Beyotime, China), anti-UBE2I antibody (Proteintech, China), anti-UBE2L3 antibody (Proteintech, China), anti-RPL15 antibody (Proteintech, China), anti-C5ORF24 antibody (Proteintech, China), and anti-FOSL2 antibody (Proteintech, China) were used for immunoblotting.

Techniques: Western Blot, Ubiquitin Proteomics, Expressing, Virus, Infection, Control, Multiplex sample analysis, Phospho-proteomics

Journal: Cell Discovery

Article Title: Structural insights into histone exchange by human SRCAP complex

doi: 10.1038/s41421-023-00640-1

Figure Lengend Snippet: a Superimposition of the two structures with differences highlighted and indicated with arrows. Nucleosomes are superimposed for comparison. Three views showing the rotation/displacement of SRCAP, hexamer, and ARP6–ZNHIT1 dimer, respectively. b – d Close-up views of the differences in ZNHIT1 ( b ) H2A C-terminal tail ( c ) and YL1 ( d ). Conformational differences are highlighted with arrows. Structural comprision showing the dissociation of ZNHIT1 from the motor-proximal H2A–H2B ( b ). Structural comparison showing the displacement of ARP6 and the possible interaction between the relocated ARP6 and the C-terminal tail of histone H2A and the sequence alignment of the C-terminal tail of H2A and H2A.Z. The subdomain4 of ARP6 is shown in electrostatic surface and H2A tails (residues 110–127) are shown in transparent surfaces and structural models ( c ). Structural comparison showing the contraction of YL1 ( d ).

Article Snippet: Antibodies were as follows: H2A.Z (ab4174, abcam), ZNHIT1 (16595-1-AP, Proteintech), VPS72 (15143-1-AP, Proteintech), YEATS4 (A6318, ABclonal), Flag (SLAB0101, Smart-Lifesciences), tubulin (AC008, Abclonal).

Techniques: Comparison, Sequencing

Journal: Cell Discovery

Article Title: Structural insights into histone exchange by human SRCAP complex

doi: 10.1038/s41421-023-00640-1

Figure Lengend Snippet: a Schematic diagram of establishing ZNHIT1-dTAG DLD-1 cells and validation of ZNHIT1 degradation and rescue. An empty vector, WT and mutant ZNHIT1 were transfected into the ZNHIT1-dTAG cells, respectively. The expression of target proteins was detected by western blot. Complex composition was validated by silver staining of the SRCAP-C purified from the indicated ZNHIT1-dTAG cells. b Metaplot and boxplot representation of H2A.Z occupancy in ZNHIT1-dTAG cells treated with DMSO or dTAG for the indicated times. c Metaplot and boxplot representation of H2A.Z occupancy in DMSO/dTAG-treated ZNHIT1-dTAG cells with overexpression of WT or mutant ZNHIT1. d Representative track examples showing H2A.Z occupancy in DMSO/dTAG-treated ZNHIT1-dTAG cells with overexpression of WT or mutant ZNHIT1. e Heatmaps of H2A.Z occupancy centered at TSSs of promoters ranked by decreasing occupancy in DMSO/dTAG-treated ZNHIT1-dTAG cells with overexpression of WT or mutant ZNHIT1.

Article Snippet: Antibodies were as follows: H2A.Z (ab4174, abcam), ZNHIT1 (16595-1-AP, Proteintech), VPS72 (15143-1-AP, Proteintech), YEATS4 (A6318, ABclonal), Flag (SLAB0101, Smart-Lifesciences), tubulin (AC008, Abclonal).

Techniques: Biomarker Discovery, Plasmid Preparation, Mutagenesis, Transfection, Expressing, Western Blot, Silver Staining, Purification, Over Expression

Journal: Cell Discovery

Article Title: Structural insights into histone exchange by human SRCAP complex

doi: 10.1038/s41421-023-00640-1

Figure Lengend Snippet: Cartoons were generated from cryo-EM maps. Right panels show close-up views of the nucleosome-bound ARP6–ZNHIT1 with other portions omitted for clarity. Multiple cycles of ATP hydrolysis may be required to destabilize H2A–H2B, followed by a successful histone exchange. Histone chaperones and associated H2A.Z–H2B may also facilitate histone exchange. One H2A–H2B is replaced in each exchange reaction.

Article Snippet: Antibodies were as follows: H2A.Z (ab4174, abcam), ZNHIT1 (16595-1-AP, Proteintech), VPS72 (15143-1-AP, Proteintech), YEATS4 (A6318, ABclonal), Flag (SLAB0101, Smart-Lifesciences), tubulin (AC008, Abclonal).

Techniques: Generated, Cryo-EM Sample Prep

Journal: Molecular and Clinical Oncology

Article Title: Stathmin1 promotes lymph node metastasis in hypopharyngeal squamous cell carcinoma via regulation of HIF‑1α/VEGF‑A axis and MTA1 expression

doi: 10.3892/mco.2023.2617

Figure Lengend Snippet: Sequences of shRNAs targeting STMN1.

Article Snippet: Equal amounts of proteins (30 μg) were added for 10% SDS-polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride membrane (Pall Life Sciences), then blocked with 5% skimmed milk at room temperature for 1.5 h. Subsequently, membranes were incubated with the following primary antibodies against: STMN1 (1:1,000; cat. no. PB9560; Wuhan Boster Biological Technology, Ltd.), hypoxia inducible factor-1alpha (HIF-1α; 1:1,000; cat. no. A6265; ABclonal Biotech Co., Ltd.), vascular endothelial growth factor (VEGF)-A (1:1,000; cat. no. ab214424; Abcam), metastasis-associated protein 1 (MTA1; 1:1,000; cat. no. ab288765; Abcam) and β-actin (1:2,000; cat. no. CPA9066; Cohesion Biosciences) overnight at 4 ̊C, and then washed with Tris-buffered saline and Tween-20 (TBST, 0.01 M Tris, 0.15 M NaCl, 0.1% Tween-20, pH=7.4) for 30 min.

Techniques: Sequencing, shRNA

Journal: Molecular and Clinical Oncology

Article Title: Stathmin1 promotes lymph node metastasis in hypopharyngeal squamous cell carcinoma via regulation of HIF‑1α/VEGF‑A axis and MTA1 expression

doi: 10.3892/mco.2023.2617

Figure Lengend Snippet: Primer sequences of genes analyzed for reverse transcription-quantitative PCR.

Article Snippet: Equal amounts of proteins (30 μg) were added for 10% SDS-polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride membrane (Pall Life Sciences), then blocked with 5% skimmed milk at room temperature for 1.5 h. Subsequently, membranes were incubated with the following primary antibodies against: STMN1 (1:1,000; cat. no. PB9560; Wuhan Boster Biological Technology, Ltd.), hypoxia inducible factor-1alpha (HIF-1α; 1:1,000; cat. no. A6265; ABclonal Biotech Co., Ltd.), vascular endothelial growth factor (VEGF)-A (1:1,000; cat. no. ab214424; Abcam), metastasis-associated protein 1 (MTA1; 1:1,000; cat. no. ab288765; Abcam) and β-actin (1:2,000; cat. no. CPA9066; Cohesion Biosciences) overnight at 4 ̊C, and then washed with Tris-buffered saline and Tween-20 (TBST, 0.01 M Tris, 0.15 M NaCl, 0.1% Tween-20, pH=7.4) for 30 min.

Techniques:

Journal: Molecular and Clinical Oncology

Article Title: Stathmin1 promotes lymph node metastasis in hypopharyngeal squamous cell carcinoma via regulation of HIF‑1α/VEGF‑A axis and MTA1 expression

doi: 10.3892/mco.2023.2617

Figure Lengend Snippet: Clinicopathological characteristics of the included cases.

Article Snippet: Equal amounts of proteins (30 μg) were added for 10% SDS-polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride membrane (Pall Life Sciences), then blocked with 5% skimmed milk at room temperature for 1.5 h. Subsequently, membranes were incubated with the following primary antibodies against: STMN1 (1:1,000; cat. no. PB9560; Wuhan Boster Biological Technology, Ltd.), hypoxia inducible factor-1alpha (HIF-1α; 1:1,000; cat. no. A6265; ABclonal Biotech Co., Ltd.), vascular endothelial growth factor (VEGF)-A (1:1,000; cat. no. ab214424; Abcam), metastasis-associated protein 1 (MTA1; 1:1,000; cat. no. ab288765; Abcam) and β-actin (1:2,000; cat. no. CPA9066; Cohesion Biosciences) overnight at 4 ̊C, and then washed with Tris-buffered saline and Tween-20 (TBST, 0.01 M Tris, 0.15 M NaCl, 0.1% Tween-20, pH=7.4) for 30 min.

Techniques: Expressing

Journal: Molecular and Clinical Oncology

Article Title: Stathmin1 promotes lymph node metastasis in hypopharyngeal squamous cell carcinoma via regulation of HIF‑1α/VEGF‑A axis and MTA1 expression

doi: 10.3892/mco.2023.2617

Figure Lengend Snippet: STMN1 expression differences and immunohistochemical staining for STMN1 and LVI. (A and B) STMN1 expression level was increased in HSCC tissues compared with normal hypopharyngeal tissues, and was further increased in HSCC tissues of patients with metastases. (C and D) STMN1 expression status in different HSCC samples (magnification, x200). (E and F) LVI statuses in different HSCC samples along with STMN1 expression statuses (magnification, x400). STMN1, stathmin1; LVI, lymphatic vessel invasion; HSCC, hypopharyngeal squamous cell carcinoma.

Article Snippet: Equal amounts of proteins (30 μg) were added for 10% SDS-polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride membrane (Pall Life Sciences), then blocked with 5% skimmed milk at room temperature for 1.5 h. Subsequently, membranes were incubated with the following primary antibodies against: STMN1 (1:1,000; cat. no. PB9560; Wuhan Boster Biological Technology, Ltd.), hypoxia inducible factor-1alpha (HIF-1α; 1:1,000; cat. no. A6265; ABclonal Biotech Co., Ltd.), vascular endothelial growth factor (VEGF)-A (1:1,000; cat. no. ab214424; Abcam), metastasis-associated protein 1 (MTA1; 1:1,000; cat. no. ab288765; Abcam) and β-actin (1:2,000; cat. no. CPA9066; Cohesion Biosciences) overnight at 4 ̊C, and then washed with Tris-buffered saline and Tween-20 (TBST, 0.01 M Tris, 0.15 M NaCl, 0.1% Tween-20, pH=7.4) for 30 min.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Molecular and Clinical Oncology

Article Title: Stathmin1 promotes lymph node metastasis in hypopharyngeal squamous cell carcinoma via regulation of HIF‑1α/VEGF‑A axis and MTA1 expression

doi: 10.3892/mco.2023.2617

Figure Lengend Snippet: The transfection efficacy evaluation of STMN1 knockdown and the results of cell functional experiments. (A and B) The relative mRNA and protein expression levels of STMN1 in different groups. All of them were evidently decreased after STMN1 knockdown with different shRNA sequences, compared with the NC shRNA group. (C-E) The OD 450 , the relative migratory ratio at 48 h, and the relative number of invasive cells were all significantly decreased after STMN1 knockdown with different shRNA sequences; all were compared with the NC shRNA group. The magnification of picture D and picture E were 100 times and 200 times, respectively. * P<0.05 and ** P<0.01. STMN1, stathmin1; shRNA, short hairpin RNA; NC, negative control.

Article Snippet: Equal amounts of proteins (30 μg) were added for 10% SDS-polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride membrane (Pall Life Sciences), then blocked with 5% skimmed milk at room temperature for 1.5 h. Subsequently, membranes were incubated with the following primary antibodies against: STMN1 (1:1,000; cat. no. PB9560; Wuhan Boster Biological Technology, Ltd.), hypoxia inducible factor-1alpha (HIF-1α; 1:1,000; cat. no. A6265; ABclonal Biotech Co., Ltd.), vascular endothelial growth factor (VEGF)-A (1:1,000; cat. no. ab214424; Abcam), metastasis-associated protein 1 (MTA1; 1:1,000; cat. no. ab288765; Abcam) and β-actin (1:2,000; cat. no. CPA9066; Cohesion Biosciences) overnight at 4 ̊C, and then washed with Tris-buffered saline and Tween-20 (TBST, 0.01 M Tris, 0.15 M NaCl, 0.1% Tween-20, pH=7.4) for 30 min.

Techniques: Transfection, Knockdown, Functional Assay, Expressing, shRNA, Negative Control

Journal: Molecular and Clinical Oncology

Article Title: Stathmin1 promotes lymph node metastasis in hypopharyngeal squamous cell carcinoma via regulation of HIF‑1α/VEGF‑A axis and MTA1 expression

doi: 10.3892/mco.2023.2617

Figure Lengend Snippet: Bioinformatics analyses for target genes and pathways of STMN1. (A) STMN1 was evidently enriched in the HIF-1α pathway in head and neck squamous cell carcinoma. (B) Gene expression levels of HIF-1α pathway were different between cases with high expression of STMN1 and those with low expression of STMN1. (C and D) Target gene prediction analysis from website https://www.aclbi.com , which demonstrated that the expression level of MTA1 was significantly correlated with that of STMN1. STMN1, stathmin1; HIF-1α, hypoxia inducible factor-1alpha; MTA1, tumor metastasis-associated protein 1.

Article Snippet: Equal amounts of proteins (30 μg) were added for 10% SDS-polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride membrane (Pall Life Sciences), then blocked with 5% skimmed milk at room temperature for 1.5 h. Subsequently, membranes were incubated with the following primary antibodies against: STMN1 (1:1,000; cat. no. PB9560; Wuhan Boster Biological Technology, Ltd.), hypoxia inducible factor-1alpha (HIF-1α; 1:1,000; cat. no. A6265; ABclonal Biotech Co., Ltd.), vascular endothelial growth factor (VEGF)-A (1:1,000; cat. no. ab214424; Abcam), metastasis-associated protein 1 (MTA1; 1:1,000; cat. no. ab288765; Abcam) and β-actin (1:2,000; cat. no. CPA9066; Cohesion Biosciences) overnight at 4 ̊C, and then washed with Tris-buffered saline and Tween-20 (TBST, 0.01 M Tris, 0.15 M NaCl, 0.1% Tween-20, pH=7.4) for 30 min.

Techniques: Gene Expression, Expressing

Journal: Molecular and Clinical Oncology

Article Title: Stathmin1 promotes lymph node metastasis in hypopharyngeal squamous cell carcinoma via regulation of HIF‑1α/VEGF‑A axis and MTA1 expression

doi: 10.3892/mco.2023.2617

Figure Lengend Snippet: The mRNA and protein expression levels of the target genes of STMN1 in FaDu cells. (A-C) The relative mRNA and protein expression levels of HIF-1α were signifiicantly decreased after STMN1 knockdown with different shRNA sequences. (D) The relative expression level of VEGF-A, the downstream protein of HIF-1α, was significantly decreased after STMN1 knockdown. (E and F) Both relative mRNA and protein expression levels of MTA1 significantly decreased after STMN1 knockdown. All were compared with the NC shRNA group. * P<0.05, ** P<0.01 and *** P<0.001. STMN1, stathmin1; HIF-1α, hypoxia inducible factor-1alpha; shRNA, short hairpin RNA; VEGF, vascular endothelial growth factor; MTA1, tumor metastasis-associated protein 1; NC, negative control.

Article Snippet: Equal amounts of proteins (30 μg) were added for 10% SDS-polyacrylamide gel electrophoresis and then transferred to polyvinylidene difluoride membrane (Pall Life Sciences), then blocked with 5% skimmed milk at room temperature for 1.5 h. Subsequently, membranes were incubated with the following primary antibodies against: STMN1 (1:1,000; cat. no. PB9560; Wuhan Boster Biological Technology, Ltd.), hypoxia inducible factor-1alpha (HIF-1α; 1:1,000; cat. no. A6265; ABclonal Biotech Co., Ltd.), vascular endothelial growth factor (VEGF)-A (1:1,000; cat. no. ab214424; Abcam), metastasis-associated protein 1 (MTA1; 1:1,000; cat. no. ab288765; Abcam) and β-actin (1:2,000; cat. no. CPA9066; Cohesion Biosciences) overnight at 4 ̊C, and then washed with Tris-buffered saline and Tween-20 (TBST, 0.01 M Tris, 0.15 M NaCl, 0.1% Tween-20, pH=7.4) for 30 min.

Techniques: Expressing, Knockdown, shRNA, Negative Control

Journal: bioRxiv

Article Title: Vector analysis of steerable mechanical tension across nuclear lamina

doi: 10.1101/462275

Figure Lengend Snippet: (A) MCF-7 cells were treated with ADM, H 2 O 2 , and CHX separately. The samples were stained for both β-actin and α-tubulin. The cells were also stained with DAPI to localize the nuclei. The images were captured through confocal laser microscopy after immunofluorescence staining (n = 10). (B) Intracellular nanoparticle size distribution in MCF-7 cells separately treated with ADM, H 2 O 2 , and CHX (n = 10). (C) MCF-7 cell intracellular osmotic pressure was measured via osmometery after the cells were exposed to ADM, H 2 O 2 , and CHX. (**: 0.001 < p < 0.05, ns: p > 0.05, Tukey-b test, n = 10). (D) P-cofilin, cofilin, actin, p-stathmin, stathmin, and tubulin levels in MCF-7 cells that had been treated with ADM, H 2 O 2 , or CHX (n = 6). (E) In the H 2 O 2 group, the ΔpNOP could be divided into two parts by comparing the total intensity of the protein nanoparticles, generated from MF and MT depolymerization, respectively (versus , line 1, row 2). Scale bar, 10 μm. All error bars represent SEM.

Article Snippet: Antibodies were purchased from commercial sources: rabbit anti-lamin B1 antibody (13435S, Cell Signaling Technology, Temecula, USA), rabbit anti-β-actin antibody (4970P, Cell Signaling Technology, Temecula, USA), mouse anti-tubulin-α antibody (T5168, Boster, Wuhan, China), rabbit anti-vimentin antibody (5741S, Cell Signaling Technology, Temecula, USA), rabbit anti-GFP antibody (2956S, Cell Signaling Technology, Temecula, USA), mouse anti-phospho-cofilin (Ser24) (bs-10252R, Bioss, Woburn, USA), mouse anti-cofilin antibody (PB0035, Boster, Wuhan, China), rabbit anti-phospho-stathmin 1 (Ser38) (bs-3432R, Bioss, Woburn, USA), and mouse anti-stathmin 1 antibody (BA1417, Boster, Wuhan, China).

Techniques: Staining, Microscopy, Immunofluorescence, Generated

Journal: Molecules

Article Title: The Large Molecular Weight Polysaccharide from Wild Cordyceps and Its Antitumor Activity on H22 Tumor-Bearing Mice

doi: 10.3390/molecules28083351

Figure Lengend Snippet: WCP promotes Cyto-c/Caspase8/3 and inhibits IL-10/STAT3/Bcl2 pathway. ( A – F ) Relative mRNA expression of IL-6, IL-Iβ, NF-κB, TNF-α, Bax, and Bcl2. * p < 0.05 compared to model group.

Article Snippet: Rabbit monoclonal antibodies against Caspase8 (PTM-6085) antibody were purchased from PTM Biolab Co., Ltd. (Hangzhou, China), and signal transducer and activator of transcription3 (Stat3, BM4052), phosphorylated signal transducer and activator of transcription3 (p-STAT3 Y705, BM4835) were purchased from Boster Biological Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing

Journal: Molecules

Article Title: The Large Molecular Weight Polysaccharide from Wild Cordyceps and Its Antitumor Activity on H22 Tumor-Bearing Mice

doi: 10.3390/molecules28083351

Figure Lengend Snippet: WCP promotes Cyto-c/Caspase8/3 and inhibits IL-10/STAT3/Bcl2pathway. ( A , F ) Photographs of the proteins in each group. ( B – E , G – I ) Relative protein expression of Cyto-c, Caspase8, Caspase3, p-STAT3, Bcl2, Bax, and Bax/Bcl2. * p < 0.05, ** p < 0.01, *** p < 0.001 compared to model group.

Article Snippet: Rabbit monoclonal antibodies against Caspase8 (PTM-6085) antibody were purchased from PTM Biolab Co., Ltd. (Hangzhou, China), and signal transducer and activator of transcription3 (Stat3, BM4052), phosphorylated signal transducer and activator of transcription3 (p-STAT3 Y705, BM4835) were purchased from Boster Biological Technology Co., Ltd. (Wuhan, China).

Techniques: Expressing