Journal: Stem Cell Research & Therapy

Article Title: Development, characterization, and hematopoietic differentiation of Griscelli syndrome type 2 induced pluripotent stem cells

doi: 10.1186/s13287-021-02364-z

Figure Lengend Snippet: Immunophenotype of BM-MNC- and BM-MSC-derived iPSC clones. a Top lane: representative histograms of a GS-2 iPSC clone (YF #D2B2) derived from BM-MNCs. Unstained controls are given as black lines and stained iPSC as red lines. Lower lane: representative histograms and dot plots from a BM-MSC-derived GS-2 iPSC clone (YF #A2C3). b Healthy donor iPSc (top) and GS-2 iPSC (bottom) immunofluorescence images, representative samples. Both healthy donor and GS-2 iPSCs show bright expression of pluripotency markers. The nuclei were counterstained with DAPI. c Relative gene expression levels of SOX2 , NANOG , and OCT4 normalized to B2M expression. Healthy (GP) and GS-2 (IK, YF, YKÇ) iPSC clones showed increased levels of SOX2 , NANOG , OCT4 gene expression in comparison with healthy donor (LA) and GS-2 (YKÇ) BM-MSCs (negative controls)

Article Snippet: Cells were blocked with a blocking buffer consisting of PBN, 1% AB serum, and 1% mouse serum. iPSCs were stained with the following primary antibodies: rabbit-anti-human OCT3/4 (Molecular Probes, A24867), mouse-anti-human SSEA4 (Molecular Probes, A24866), mouse-anti-human TRA-1-60 (Molecular Probes, A24868), rabbit-anti-OCT4A (Cell Signaling Technology, 2840), rat-anti-SOX2 (Cell Signaling Technology, 3579), rabbit-anti-NANOG (Cell Signaling Technology, 4903), rabbit-anti-c-MYC (Cell Signaling Technology, 5605), and rabbit-anti-LIN28A (Cell Signaling Technology, 3695).

Techniques: Derivative Assay, Clone Assay, Staining, Immunofluorescence, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Membrane Transporters in Human Parotid Gland-Targeted Proteomics Approach

doi: 10.3390/ijms20194825

Figure Lengend Snippet: Correlation between mRNA and protein level of ABC and SLC transporters in the human parotid salivary gland. Correlation coefficients were assessed using the Spearman’s rank test. Bold font indicates p value < 0.05.

Article Snippet: After serum incubation, the slides were incubated with primary antibodies: rabbit polyclonal anti-human OCT3 (Sigma Aldrich, Taufkirchen, Germany), rabbit polyclonal anti-human MATE1 (Sigma Aldrich, Taufkirchen, Germany), mouse monoclonal anti-human MDR1 (Sigma Aldrich, Taufkirchen, Germany) and mouse monoclonal anti-human MRP1 (Abcam, Cambridge, UK) for 1 hour in RT, and after double washing in PBS, the slides were incubated with goat anti rabbit AlexaFluor 488 (Life Technologies, Waltham, MA, USA) for 1 hour at RT in the dark.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Membrane Transporters in Human Parotid Gland-Targeted Proteomics Approach

doi: 10.3390/ijms20194825

Figure Lengend Snippet: Immunofluorescence staining of OCT3 and MATE1 in salivary glands and kidney cortex. ( A ) OCT3 (green panel) expression at the basolateral and apical membrane of serous and mucous acinar cells and in duct cells. ( B ) OCT3 (green panel) expression at basolateral membrane of kidney proximal tubule. ( C ) MATE1 (green panel) detection at the basolateral and apical membrane of serous and mucous acinar cells and in duct cells. ( D ) MATE1 (green panel) detection at apical membrane of kidney proximal tubule. Na + /K + -ATPase (red panel A – D ) was used as the basolateral membrane marker. Nuclei are stained blue ( A – D ). Yellow arrow—apical and basolateral membrane of mucous acinar cells; white arrow—apical and basolateral membrane of duct cells; pink arrow—serous acinar cells; blue and red arrow—basolateral and apical membrane of proximal tubule cells, respectively.

Article Snippet: After serum incubation, the slides were incubated with primary antibodies: rabbit polyclonal anti-human OCT3 (Sigma Aldrich, Taufkirchen, Germany), rabbit polyclonal anti-human MATE1 (Sigma Aldrich, Taufkirchen, Germany), mouse monoclonal anti-human MDR1 (Sigma Aldrich, Taufkirchen, Germany) and mouse monoclonal anti-human MRP1 (Abcam, Cambridge, UK) for 1 hour in RT, and after double washing in PBS, the slides were incubated with goat anti rabbit AlexaFluor 488 (Life Technologies, Waltham, MA, USA) for 1 hour at RT in the dark.

Techniques: Immunofluorescence, Staining, Expressing, Marker

Journal: PLoS ONE

Article Title: MicroRNAs Induce Epigenetic Reprogramming and Suppress Malignant Phenotypes of Human Colon Cancer Cells

doi: 10.1371/journal.pone.0127119

Figure Lengend Snippet: A) Scheme of cancer reprogramming methods by transfection of miR-302s or miR-302s plus miR-369s. B) miR-302s-induced morphological changes in HT29 cells at days 1, 6, and 28. Reprogrammed HT29 have an embryonic stem cells-like appearance. Scale bars, 100 μm. C) Cells stained using alkaline phosphatase (AP) live stain. Scale bars, 100 μm. D) Relative expression of OCT3/4 , SOX2 and NANOG compared to AP − cells by qRT-PCR (n = 3). E) Relative expression of miR-302s and miR-369s in AP + , AP − , and NTERA cells. Mean expression of each miR was compared to that in NTERA cells (n = 3). F) Immunofluorescence of pluripotent stem cell markers Oct3/4, Sox2, and Nanog. Nuclei were stained with DAPI. Scale bars, 100 μm (original magnification, ×200).

Article Snippet: Slides were heated in antigen retrieval buffer for 40 min, blocked with goat or horse serum for 20 min at room temperature, and incubated with monoclonal mouse anti-human Ki67 antigen (1:400, DAKO), polyclonal rabbit anti-human Oct3/4 (1:100, MBL), polyclonal rabbit anti-human Sox2 (1:200, MBL), or monoclonal mouse anti-human CK20 (DAKO) antibodies overnight at 4°C.

Techniques: Transfection, Staining, Expressing, Quantitative RT-PCR, Immunofluorescence

Journal: PLoS ONE

Article Title: MicroRNAs Induce Epigenetic Reprogramming and Suppress Malignant Phenotypes of Human Colon Cancer Cells

doi: 10.1371/journal.pone.0127119

Figure Lengend Snippet: A) Tumor-bearing mice were treated with systemic administration of carbonate apatite (CA)- complexed miR-302s, negative control (NC) miR, or CA only every second day, with a total of six injections. Tumor growth curves are shown. Asterisk denotes a p-value in the Mann-Whitney U-test of < 0.05. B) Tumor-bearing mice were treated with systemic administration of CA-equipped miR-302s plus miR-369s, NC miR, or CA only daily, with a total of 14 injections, similar to (A). C) Representative hematoxylin and eosin-stained images of tumor sections from the indicated experimental groups. Areas with denaturated nuclei were predominantly observed in xenografts treated with miR-302s/CA complexes and miR-302s plus miR-369s/CA complexes. Scale bars, 100 μm (magnification ×200). D) Confirmation of intratumoral apoptosis by TUNEL staining in tumor sections obtained from xenografts treated with CA-mediated miR-302s, miR-302s plus miR-369s, NC, or CA only. Scale bars, 100 μm (magnification ×200). E) Immunohistochemistry analysis of the proliferation marker Ki-67 in xenografts treated with miR-302s, miR-302s plus miR-369s, NC miR or the mock control. Representative images are shown. Scale bars, 100 μm (magnification ×200). Ki-67-positive cells were counted in three random fields from different three xenografts in each group. F) Relative expression of pluripotency-related markers OCT3/4, SOX2 and NANOG in the indicated experimental groups. Data are shown in comparison with the NC miR group. (n = 3). G) Representative immunohistochemical images of tumor sections from the indicated experiment groups. Oct3/4 and Sox2 were predominantly observed in xenografts treated with miR-302s and miR-302s plus miR-369s. Alcian blue staining showed the decrease of mucin production in these miR-treated-groups. CK20 expression was also decreased in xenografts treated with miR-302s plus miR-369s. Scale bars, 100 μm (magnification ×200).

Article Snippet: Slides were heated in antigen retrieval buffer for 40 min, blocked with goat or horse serum for 20 min at room temperature, and incubated with monoclonal mouse anti-human Ki67 antigen (1:400, DAKO), polyclonal rabbit anti-human Oct3/4 (1:100, MBL), polyclonal rabbit anti-human Sox2 (1:200, MBL), or monoclonal mouse anti-human CK20 (DAKO) antibodies overnight at 4°C.

Techniques: Negative Control, MANN-WHITNEY, Staining, TUNEL Assay, Immunohistochemistry, Marker, Expressing, Immunohistochemical staining