Journal: Stem Cell Research & Therapy

Article Title: hAMSCs regulate EMT in the progression of experimental pulmonary fibrosis through delivering miR-181a-5p targeting TGFBR1

doi: 10.1186/s13287-024-04095-3

Figure Lengend Snippet:

Article Snippet: Anti-Collagen Type I/COL1A1 Antibody , BOSTER BA0325.

Techniques: Isolation, Reverse Transcription, Fluorescence, Real-time Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Microvesicles from Mesenchymal Stromal Cells Are Involved in HPC-Microenvironment Crosstalk in Myelodysplastic Patients

doi: 10.1371/journal.pone.0146722

Figure Lengend Snippet: (A) Percentage of CD34 + cells isolated by immunomagnetic beads and the purity determined by flow cytometry. (B) Flow cytometry characterization of MVs released from MSCs of MDS and HD. The upper images are dot-plots of forward and side scatter of MVs. The gate was defined as elements of smaller size than the 1μm beads. The histograms represent the MVs stained with negative (CD34 and CD45) and positive markers for MVs from HD and MDS-MSC (CD90, CD44, CD73) and for MVs markers (CD81 and CD63). Controls (unstained MVs) are shown in gray; the MVs stained with the different antibodies are shown in black. Images on the left are those of the MVs from MSC-HD, while those on the right images are of the MVs from MSC-MDS. (C) Representative images of transmission electronic microscopy of MVs released by MSC from HD(left) and MDS (right) as revealed by TEM. Scale bar, 200nm. Original magnification: x 8000. (D) MVs characterization by Western Blot assay for the expression of CD63. HD-MVs: microvesicles from healthy donors. MDS-MVs: microvesicles from patients with myelodysplastic syndrome.

Article Snippet: After isolation, the purity and viability of CD34 + cells was evaluated by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated CD34 (11-0349-42, eBioscience Inc. San Diego, CA, USA) and APC H7 Annexin V DY634.

Techniques: Isolation, Flow Cytometry, Staining, Transmission Assay, Microscopy, Western Blot, Expressing

Journal: PLoS ONE

Article Title: Microvesicles from Mesenchymal Stromal Cells Are Involved in HPC-Microenvironment Crosstalk in Myelodysplastic Patients

doi: 10.1371/journal.pone.0146722

Figure Lengend Snippet: (A) Representative images of MVs incorporation by CD34 + cells stained with anti-CD90 Ab (red) and anti-CD45 Ab (green). (B) Representative images of MVs previously labeled with Vybrant-Dil cell-labeling solution (red) that were incorporated into CD34 + cells and stained with anti-CD45 Ab (green). (A-B) Images in the top row are from CD34 + cells that incorporated the MVs released from MSC-HD. Images on the middle row show the incorporation of MVs released from MSC-MDS. In the lower row, images of the CD34 + cells (without incorporation) are shown. Nuclei were counterstained with DAPI (blue). Scale bar, 7.5μm. Revealed by confocal microscopy and acquired in layers (z-Stacks) of 1μm.

Article Snippet: After isolation, the purity and viability of CD34 + cells was evaluated by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated CD34 (11-0349-42, eBioscience Inc. San Diego, CA, USA) and APC H7 Annexin V DY634.

Techniques: Staining, Labeling, Confocal Microscopy

Journal: PLoS ONE

Article Title: Microvesicles from Mesenchymal Stromal Cells Are Involved in HPC-Microenvironment Crosstalk in Myelodysplastic Patients

doi: 10.1371/journal.pone.0146722

Figure Lengend Snippet: (A) Variations in microRNAs expression when CD34 + cells were co-cultured with MDS-MVs or HD-MVs. Ratio was calculated dividing the expression of each microRNA from CD34 + + HD-MVs or CD34 + + MDS-MVs by that of CD34 + cells without MVs. Results were summarized as the median. (B) Expression by RT-PCR of TP53 and MDM2 in CD34 + cells cultured with MDS-MVs from patients (grey) and expression of CD34 + cells without MVs (black). Results were summarized as the mean and standard deviation. (C) Capillary Electrophoresis Immunoassay of MDM2 vs Actin as control. CD34 + cells (without MVs), CD34 + cells with MDS-MVs and with HD-MVs. Each bar of the lower graph represents the value of quantified MDM2 protein expression normalized to actin protein abundance. Each bar represents the quantification of both bands of MDM2 from the pseudo-blots, control CD34 + cells vs CD34 + cells + MDS-MVs or HD-MVs.

Article Snippet: After isolation, the purity and viability of CD34 + cells was evaluated by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated CD34 (11-0349-42, eBioscience Inc. San Diego, CA, USA) and APC H7 Annexin V DY634.

Techniques: Expressing, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Standard Deviation, Electrophoresis, Control, Quantitative Proteomics

Journal: PLoS ONE

Article Title: Microvesicles from Mesenchymal Stromal Cells Are Involved in HPC-Microenvironment Crosstalk in Myelodysplastic Patients

doi: 10.1371/journal.pone.0146722

Figure Lengend Snippet: (A): Representative FACS plots of annexinV/7AAD staining on CD34 + cells with and without MVs. Percentage of each subset (dead, live, early and late apoptosis) within the total number of CD34 + cells. (B) Percentage of live CD34 + cells. An increase on the percentage of CD34 + viable cells (annexinV - /7AAD - ) was observed when cells were cultured with MDS-MVs compared with the other groups is shown. (C) Clonogenic assays. Results are expressed as the ratio between CFU-GM obtained with CD34 + cells that had been cultured with MVs and CD34 + cells without MVs.

Article Snippet: After isolation, the purity and viability of CD34 + cells was evaluated by flow cytometry using fluorescein isothiocyanate (FITC)-conjugated CD34 (11-0349-42, eBioscience Inc. San Diego, CA, USA) and APC H7 Annexin V DY634.

Techniques: Staining, Cell Culture

Journal: Cancer biology & therapy

Article Title: CXCR4 confers stemness and radioresistance in chordoma cells.

doi: 10.1080/15384047.2025.2471631

Figure Lengend Snippet: Figure 1. CXCR4+ DTC cells exhibit enhanced self-renewal activity, tumorigenic potential and IR resistance compared to CXCR4- cells. (a) MTT assay, (b) sphere-forming assay (white bar 50 mm), (c) limiting dilution assay, (d) soft agar assay (white bar 50 μm), (e) IR cl, 22onogenic survival assay (black bar 50 mm), and (f) Western blot (left) and RT-PCR (right) analyses of DTC cells after sorting with apc-conjugated CXCR4 antibody. *p < .05. All experimental results were obtained from at least three independent experiments.

Article Snippet: To sort CXCR4+ and CXCR4− cells, DTC cells were dissociated into single cells, washed with ice-cold PBS, and stained with an APC-conjugated CXCR4 antibody (#130-100-070; Miltenyi Biotec Inc., Auburn, CA, USA) in 0.1 ml of binding buffer (2% BSA and 5 mm EDTA in PBS) at a 4°C refrigerator for 1 h. After washing with ice-cold PBS three times, cells were incubated with anti-APC microbead in 0.1 ml of binding buffer at a 4°C refrigerator for 30 min.

Techniques: Activity Assay, MTT Assay, Limiting Dilution Assay, Soft Agar Assay, Clonogenic Cell Survival Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Cancer biology & therapy

Article Title: CXCR4 confers stemness and radioresistance in chordoma cells.

doi: 10.1080/15384047.2025.2471631

Figure Lengend Snippet: Figure 2. Silencing of CXCR4 via shRNA infection suppresses self-renewal activity, tumorigenic potential and IR resistance in DTC cells. (a) MTT assay, (b) sphere-forming assay (white bar 50 mm), (c) limiting dilution assay, (d) soft agar assay (white bar 50 μm), (e) IR clonogenic survival assay (black bar 50 mm), (f) Western blot (left) and RT-PCR (right), and (g) immunofluorescence (white bar 100 μm) analyses of DTC cells infected with shControl (shCTL), shCXCR4 a or D (upper) and quantification of the results (lower). *p < .05, **p < .01, ***p < .001. All experimental results were obtained from at least three independent experiments.

Article Snippet: To sort CXCR4+ and CXCR4− cells, DTC cells were dissociated into single cells, washed with ice-cold PBS, and stained with an APC-conjugated CXCR4 antibody (#130-100-070; Miltenyi Biotec Inc., Auburn, CA, USA) in 0.1 ml of binding buffer (2% BSA and 5 mm EDTA in PBS) at a 4°C refrigerator for 1 h. After washing with ice-cold PBS three times, cells were incubated with anti-APC microbead in 0.1 ml of binding buffer at a 4°C refrigerator for 30 min.

Techniques: shRNA, Infection, Activity Assay, MTT Assay, Limiting Dilution Assay, Soft Agar Assay, Clonogenic Cell Survival Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence

Journal: Cancer biology & therapy

Article Title: CXCR4 confers stemness and radioresistance in chordoma cells.

doi: 10.1080/15384047.2025.2471631

Figure Lengend Snippet: Figure 5. CXCR4 overexpressing U-CH1 cells exhibited higher self-renewal activity, tumorigenic potential and IR resistance than control cells. (a) MTT assay, (b) sphere- forming assay (white bar 50 mm), (c) limiting dilution assay, (d) soft agar assay (white bar 50 μm), (e) IR clonogenic survival assay (black bar 50 mm), (f) Western blot (left) and RT-PCR (right), and (g) immunofluorescence (white bar 100 μm) analyses of DTC cells of transduced with control (pQCXIP) and CXCR4 carrying retroviruses in U-CH1 cells (upper) and quantification of the results (lower). *p < .05, **p < .01. All experimental results were obtained from at least three independent experiments.

Article Snippet: To sort CXCR4+ and CXCR4− cells, DTC cells were dissociated into single cells, washed with ice-cold PBS, and stained with an APC-conjugated CXCR4 antibody (#130-100-070; Miltenyi Biotec Inc., Auburn, CA, USA) in 0.1 ml of binding buffer (2% BSA and 5 mm EDTA in PBS) at a 4°C refrigerator for 1 h. After washing with ice-cold PBS three times, cells were incubated with anti-APC microbead in 0.1 ml of binding buffer at a 4°C refrigerator for 30 min.

Techniques: Activity Assay, Control, MTT Assay, Limiting Dilution Assay, Soft Agar Assay, Clonogenic Cell Survival Assay, Western Blot, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Transduction

Journal: Cancer biology & therapy

Article Title: CXCR4 confers stemness and radioresistance in chordoma cells.

doi: 10.1080/15384047.2025.2471631

Figure Lengend Snippet: Figure 6. Suppression of CXCR4 reduced tumorigenic potential and enhanced IR sensitivity of DTC cell in an in vivo xenograft model. (a) A representative image of tumor-bearing mice (left) and measurement of in vivo tumor growth rate (right) in shControl (shCTL) and shCXCR4-infected DTC cells subcutaneously transplanted in mice. (b) Combination therapy of IR and CXCR4 inhibitor (AMD3100, 5 mg/kg) in xenograft mouse model. (c) Immunohistochemistry (white bar 50 μm) of xenograft tissues probed with Sox2 antibody (left) and quantitation of the results (right) *p < .05, ***p < .001.

Article Snippet: To sort CXCR4+ and CXCR4− cells, DTC cells were dissociated into single cells, washed with ice-cold PBS, and stained with an APC-conjugated CXCR4 antibody (#130-100-070; Miltenyi Biotec Inc., Auburn, CA, USA) in 0.1 ml of binding buffer (2% BSA and 5 mm EDTA in PBS) at a 4°C refrigerator for 1 h. After washing with ice-cold PBS three times, cells were incubated with anti-APC microbead in 0.1 ml of binding buffer at a 4°C refrigerator for 30 min.

Techniques: In Vivo, Infection, Immunohistochemistry, Quantitation Assay

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

Article Title: SAMSN1 Is a Tumor Suppressor Gene in Multiple Myeloma 1 2

doi: 10.1016/j.neo.2014.07.002

Figure Lengend Snippet: SAMSN1 expression is reduced in CD138 + PCs of patients with MM and HMCL. (A) SAMSN1 expression (as determined by real-time PCR) is significantly reduced in the BMs of patients with MM ( n = 34) compared with patients with MGUS ( n = 9) and healthy age-matched controls ( n = 5; * P < .05, ** P < .001, one-way ANOVA with Tukey’s multiple comparison test). (B) SAMSN1 expression in CD138 + MACS isolated PCs from patients with MM negatively correlates with BM PC burden ( n = 10, r 2 = 0.6147, P = .0043). (C) In silico analysis of published microarray data. CD138 + PCs were isolated by MACS from 414 patients with MM, 44 patients with MGUS, and 22 age-matched controls. RNA was extracted and analyzed using the Affymetrix U133Plus2.0 microarray platform (GEO Accession Nos GSE4581 and GSE5900). Expression of SAMSN1 is significantly reduced in PCs of patients with MM compared to those of patients with MGUS and normal controls. P < 0.0001, one-way ANOVA with Tukey’s multiple comparison test. (D) Total RNA was extracted from six HMCLs and reverse transcribed. The levels of SAMSN1 expression were assessed by real-time PCR.

Article Snippet: PCs were isolated from flushed long bones and identified using rat anti-mouse CD138 (R&D Systems, Minneapolis, MN) followed by PE-conjugated goat anti-rat IgG.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Comparison, Isolation, In Silico, Microarray, Reverse Transcription

Journal: Oncogene

Article Title: Epigenetic inactivation of class II transactivator (CIITA) is associated with the absence of interferon-gamma-induced HLA-DR expression in colorectal and gastric cancer cells.

doi: 10.1038/sj.onc.1208144

Figure Lengend Snippet: Figure 1 Expression of HLA-DR in colorectal and gastric cancer cell lines. (a) RT–PCR analyses of HLA-DR expression. cDNAs were prepared from cells treated with either IFN-g or mock. Integrity of the mRNA was confirmed by amplifying GAPDH. Corresponding negative controls (amplification without reverse transcription) are shown as RT-negative. (b) Representative cell surface expression of HLA-DR protein in gastric cancer cell lines. Cell lines were treated for 48 h with either mock or 100 U/ml IFN-g, after which HLA-DR expression was examined by flow cytometry

Article Snippet: After treating the cell lines with 100 U/ml IFN-g for 48 h, they were incubated first with fluorescein isothiocyanate (FITC)conjugated anti-HLA-DR antibody (L243, ATCC), washed, and then incubated with affinity-purified, fluorescein-labeled goat anti-mouse IgGþ IgM secondary antibody (Kirkegaard & Perry Laboratories, Gaithersburg, MD, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Cytometry

Journal: Oncogene

Article Title: Epigenetic inactivation of class II transactivator (CIITA) is associated with the absence of interferon-gamma-induced HLA-DR expression in colorectal and gastric cancer cells.

doi: 10.1038/sj.onc.1208144

Figure Lengend Snippet: Figure 3 Expression of HLA-DR is associated with expression of CIITA-PIV in colorectal and gastric cancer cells. (a) Schematic representation of the structure of the CIITA gene. Three sets of primers (PI-F/R, PIII-F/R and PIV-F/R) were used to separately amplify the three CIITA isoforms. The location of the primers are indicated by the arrow. (b) Expression of CIITA-PIV, RFX5 and RFXAP in colorectal and gastric cancer cells. cDNA were prepared from cells treated with either IFN-g or mock; the integrity of mRNA was determined by amplifying GAPDH. The cell lines used are shown on the top. (c) Induction of HLA-DR by exogenous CIITA-PVI: RT–PCR analysis of CIITA-PIV and HLA-DR mRNA expression. (d) Immunofluorescence images showing expression of HLA-DR protein with (CIITA) and without (mock) introduction of exogenous CIITA-PVI. RKO cells, which do not express CIITA, were transfected with CIITA-PIV, after which expression of HLA-DR protein was analysed by immunofluorescence microscopy. The nuclei were stained with DAPI

Article Snippet: After treating the cell lines with 100 U/ml IFN-g for 48 h, they were incubated first with fluorescein isothiocyanate (FITC)conjugated anti-HLA-DR antibody (L243, ATCC), washed, and then incubated with affinity-purified, fluorescein-labeled goat anti-mouse IgGþ IgM secondary antibody (Kirkegaard & Perry Laboratories, Gaithersburg, MD, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Microscopy, Staining

Journal: Oncogene

Article Title: Epigenetic inactivation of class II transactivator (CIITA) is associated with the absence of interferon-gamma-induced HLA-DR expression in colorectal and gastric cancer cells.

doi: 10.1038/sj.onc.1208144

Figure Lengend Snippet: Figure 8 The role of DNMT1 and DNMT3B in the silencing of CIITA-PIV gene expression. (a) Analysis of CIITA methylation in DNMT1/, DNMT3B/ and DNMT1/DNMT3B/ (DKO) cells. The methylation status of CIITA was examined using primers CIITAGM1 and CIITAseqR. PCR products were digested with BstUI and then electrophoresed; M, methylated alleles. Percentages of methylated alleles are shown below the column. (b) Bisulfite-sequencing of CIITA-PIV. Bisulfite-PCR was carried out using primers GM1F and SeqR, after which direct sequencing was carried out to examine methylation. The cell lines used are shown below; closed circles, methylated cytosines; open circles, unmethylated cytosines. (c) Expression of CIITA-PIV, HLA-DR, HPP1/TPEF and COL1A in HCT116 and DKO cells. Expression of CIITA-PIV was examined with (IFN-g) and without (mock) IFN-g treatment. (d) Effect of 5-aza-dC on expression of CIITA-PIV and HLA-DR. Caco2 and HCT116 cells were treated with mock or 1 mM 5-aza-dC for 96 h and then treated with 100 U of IFN-g for 48 h

Article Snippet: After treating the cell lines with 100 U/ml IFN-g for 48 h, they were incubated first with fluorescein isothiocyanate (FITC)conjugated anti-HLA-DR antibody (L243, ATCC), washed, and then incubated with affinity-purified, fluorescein-labeled goat anti-mouse IgGþ IgM secondary antibody (Kirkegaard & Perry Laboratories, Gaithersburg, MD, USA).

Techniques: Gene Expression, Methylation, Sequencing, Expressing

Journal: Oncogene

Article Title: Epigenetic inactivation of class II transactivator (CIITA) is associated with the absence of interferon-gamma-induced HLA-DR expression in colorectal and gastric cancer cells.

doi: 10.1038/sj.onc.1208144

Figure Lengend Snippet: Figure 9 Immunohistochemical analysis of HLA-DR in primary gastric cancers. Representative images show immunofluorescent staining of HLA-DR in primary gastric cancers. Tissue sections were stained with anti-HLA-DR (green); nuclei were stained with DAPI (blue). The cases examined are indicated on the left

Article Snippet: After treating the cell lines with 100 U/ml IFN-g for 48 h, they were incubated first with fluorescein isothiocyanate (FITC)conjugated anti-HLA-DR antibody (L243, ATCC), washed, and then incubated with affinity-purified, fluorescein-labeled goat anti-mouse IgGþ IgM secondary antibody (Kirkegaard & Perry Laboratories, Gaithersburg, MD, USA).

Techniques: Immunohistochemical staining, Staining