Journal: Journal of Translational Medicine

Article Title: LncRNA H19-rich extracellular vesicles derived from gastric cancer stem cells facilitate tumorigenicity and metastasis via mediating intratumor communication network

doi: 10.1186/s12967-023-04055-0

Figure Lengend Snippet: H19 delivered by CSCs-EVs induces YAP activation and augments CDX2 expression in NSCCs. A Expression of active YAP in the nucleus of NSCCs co-cultured with CSCs-EVs or CSCs-EVs-sh-H19 detected by immunofluorescence staining. B Expression of YAP and CDX2 in the nucleus of NSCCs treated with Verteporfin detected by immunofluorescence staining. C mRNA expression of CDX2 in NSCCs treated with Verteporfin determined by RT-qPCR. D H19 and CDX2 expression in NSCCs treated with CSCs-EVs + DMSO or CSCs-EVs + Verteporfin detected by RT-qPCR. E Expression of active YAP and CDX2 in the nucleus of NSCCs treated with CSCs-EVs + DMSO or CSCs-EVs + Verteporfin detected by immunofluorescence staining. F Western blots and quantification analysis of nuclear YAP, total YAP and CDX2 proteins in NSCCs treated with CSCs-EVs + DMSO or CSCs-EVs + Verteporfin. The grayscale analysis of the relative nuclear YAP expression used H3 as the internal reference, and that of the total YAP protein expression and CDX2 protein expression used β-actin as the internal reference. Cell experiments were conducted three times independently. * p < 0.05, ** p < 0.01

Article Snippet: The sections were blocked with normal goat serum and immunostained with primary antibodies to active YAP, CDX2 and Rab27a overnight at 4 oC, and then with biotin-labeled secondary antibody goat anti-rabbit or goat anti-mouse (BM3894 or BM3895, 1: 500, Boster).

Techniques: Activation Assay, Expressing, Cell Culture, Immunofluorescence, Staining, Quantitative RT-PCR, Western Blot

Journal: Journal of Translational Medicine

Article Title: LncRNA H19-rich extracellular vesicles derived from gastric cancer stem cells facilitate tumorigenicity and metastasis via mediating intratumor communication network

doi: 10.1186/s12967-023-04055-0

Figure Lengend Snippet: H19 delivered by CSCs-EVs facilitates the malignant behaviors of NSCCs by disrupting the YAP/CDX2 signaling axis. A mRNA expression of CDX2 in NSCCs treated with oe-CDX2 detected by RT-qPCR. B Western blot of CDX2 protein in NSCCs treated with oe-CDX2. C H19 expression and CDX2 mRNA expression in NSCCs analyzed by RT-qPCR. D Western blots and quantification analysis of the total YAP and nuclear YAP proteins and CDX2 protein in the NSCCs co-cultured with CSCs-EVs-sh-H19 + oe-NC or CSCs-EVs-sh-H19 + oe-CDX2. The grayscale analysis of the relative nuclear YAP expression used H3 as the internal reference, and that of the total YAP protein expression and CDX2 protein expression used β-actin as the internal reference. E Viability of NSCCs co-cultured with CSCs-EVs-sh-H19 + oe-NC or CSCs-EVs-sh-H19 + oe-CDX2 detected by CCK-8 assay. F Migration and invasion of NSCCs co-cultured with CSCs-EVs-sh-H19 + oe-NC or CSCs-EVs-sh-H19 + oe-CDX2 detected by Transwell assay. G Western blot of the stemness marker proteins CD44 and CD133 in the NSCCs co-cultured with CSCs-EVs-sh-H19 + oe-NC or CSCs-EVs-sh-H19 + oe-CDX2. Cell experiments were conducted three times independently. * p < 0.05, ** p < 0.01

Article Snippet: The sections were blocked with normal goat serum and immunostained with primary antibodies to active YAP, CDX2 and Rab27a overnight at 4 oC, and then with biotin-labeled secondary antibody goat anti-rabbit or goat anti-mouse (BM3894 or BM3895, 1: 500, Boster).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Cell Culture, CCK-8 Assay, Migration, Transwell Assay, Marker

Journal: Journal of Translational Medicine

Article Title: LncRNA H19-rich extracellular vesicles derived from gastric cancer stem cells facilitate tumorigenicity and metastasis via mediating intratumor communication network

doi: 10.1186/s12967-023-04055-0

Figure Lengend Snippet: H19 delivered by CSCs-EVs activates the YAP/CDX2 signaling axis and facilitates the tumorigenicity and metastasis of NSCCs in vivo. A H19 and CDX2 expression in tumor tissues of mice treated with CSCs-EVs or CSCs-EVs-sh-H19 analyzed by RT-qPCR. B Positive expression of CDX2 and active YAP proteins in tumor tissues of mice treated with CSCs-EVs or CSCs-EVs-sh-H19 detected by immunohistochemistry. C Tumor weight of mice treated with CSCs-EVs or CSCs-EVs-sh-H19. D, Representative images of liver tissues and the quantitative analysis of liver metastases in mice treated with CSCs-EVs or CSCs-EVs-sh-H19. E, H&E staining of tumor tissues and liver tissues of mice treated with CSCs-EVs or CSCs-EVs-sh-H19. * p < 0.05. n = 8 mice for each treatment

Article Snippet: The sections were blocked with normal goat serum and immunostained with primary antibodies to active YAP, CDX2 and Rab27a overnight at 4 oC, and then with biotin-labeled secondary antibody goat anti-rabbit or goat anti-mouse (BM3894 or BM3895, 1: 500, Boster).

Techniques: In Vivo, Expressing, Quantitative RT-PCR, Immunohistochemistry, Staining

Journal: Journal of Translational Medicine

Article Title: LncRNA H19-rich extracellular vesicles derived from gastric cancer stem cells facilitate tumorigenicity and metastasis via mediating intratumor communication network

doi: 10.1186/s12967-023-04055-0

Figure Lengend Snippet: Schematic representation summarizing the role of CSCs-EVs carrying H19 in GC progression. CSCs-EVs can deliver H19 into NSCCs, where H19 activates the YAP/CDX2 signaling axis and facilitates the malignant phenotypes of NSCCs, ultimately promoting GC progression

Article Snippet: The sections were blocked with normal goat serum and immunostained with primary antibodies to active YAP, CDX2 and Rab27a overnight at 4 oC, and then with biotin-labeled secondary antibody goat anti-rabbit or goat anti-mouse (BM3894 or BM3895, 1: 500, Boster).

Techniques:

Journal: Turkish journal of biology = Turk biyoloji dergisi

Article Title: Correction of Griscelli Syndrome Type 2 causing mutations in the RAB27A gene with CRISPR/Cas9.

doi: 10.55730/1300-0152.2705

Figure Lengend Snippet: Figure 2. RAB27A expression by donor and GS-2 MSCs and iPSCs. Donor and GS-2 patient-derived MSCs (IK and YF) were stained for RAB27A expression using flowcytometry (upper left) and immunofluorescent staining of adherent cells in culture (upper right, DAPI: blue, RAB27A: red). RAB27A gene expression in GS-2 MSCs from two donors (İK and YF) was calculated relative to RAB27A expression in healthy donor MSCs, whereas GS-2 iPSCs expression of RAB27A was calculated relative to healthy donor iPSCs (lower left). GS-2 iPSC expression of pluripotency genes was confirmed for OCT4, SOX2, and NANOG (lower right).

Article Snippet: Membranes were stained with an anti-RAB27A antibody (#STJ25258, St John’s Laboratory Ltd, London, UK) at 1:1000 overnight at 4 °C and counterstained with secondary antirabbit-HRP antibody at 1:10000 (#11-4220-82, Invitrogen, Waltham, MA, USA) for 1 h at room temperature.

Techniques: Expressing, Derivative Assay, Staining, Gene Expression

Journal: Turkish journal of biology = Turk biyoloji dergisi

Article Title: Correction of Griscelli Syndrome Type 2 causing mutations in the RAB27A gene with CRISPR/Cas9.

doi: 10.55730/1300-0152.2705

Figure Lengend Snippet: Figure 5. Transfection of GS-2 iPSCs with the RAB27A exon 3 mutation (upper left) resulted in loss of viability and spontaneous differentiation (lower left) and low HDR efficacy (right).

Article Snippet: Membranes were stained with an anti-RAB27A antibody (#STJ25258, St John’s Laboratory Ltd, London, UK) at 1:1000 overnight at 4 °C and counterstained with secondary antirabbit-HRP antibody at 1:10000 (#11-4220-82, Invitrogen, Waltham, MA, USA) for 1 h at room temperature.

Techniques: Transfection, Mutagenesis

Journal: Particle and Fibre Toxicology

Article Title: Iron oxide nanoparticles suppress the production of IL-1beta via the secretory lysosomal pathway in murine microglial cells

doi: 10.1186/1743-8977-10-46

Figure Lengend Snippet: IONPs attenuated the cathepsin B enzyme activity of secretory lysosomes in LPS-stimulated microglia. Primary microglial cells (4 × 10 5 cells/mL) were either left untreated (naïve; NA), or pretreated with IONPs (1–50 μg Fe/mL) for 30 min followed by stimulation with LPS (100 ng/mL) for 24 h. (A) The cells pretreated with IONPs and stimulated with LPS were incubated with the cathepsin B substrate (red) for 1 h at 37°C and then stained for the secretory lysosome marker Rab27a (green). The fluorescence was visualized by confocal microscopy. Note the presence of orange signals in the merged images indicating the colocalization of cathepsin B and Rab27a. (B) The red fluorescence of 5000 single cells was measured by flow cytometry. Data are expressed as the mean ± SE of triplicate cultures. Results are a representative of three independent experiments. * p < 0.05 compared to the control group. MFI, mean fluorescence intensity.

Article Snippet: The secretory lysosomes in microglia were stained with appropriately diluted Alexa Fluor 488-labeled rabbit anti-Rab27a antibody (Bioss, MA) and visualized by confocal microscopy as described above for ED-1 staining.

Techniques: Activity Assay, Incubation, Staining, Marker, Fluorescence, Confocal Microscopy, Flow Cytometry