Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: SC transplantation shifted the CD11b immune cell population from an Arg1 − iNOS + pro-inflammatory to an intermediate Arg1 + iNOS + phenotype after SCI. Representative images of flow cytometry analysis and pie charts of CD11b population dynamics at 14 days post-injury (7 days post-transplantation) show, compared with SCI controls ( A – C ), a decreased percentage of CD11b cells stained with Arg1 − iNOS + and an increased percentage for Arg1 + iNOS + in animals receiving SC transplants ( D – F ). Results are expressed as mean ± standard deviation (SD). Abbreviations on the graphs are: Fluorescein isothiocyanate (FITC), Allophycocyanin (APC) and Forward Scatter (FSC-A). For panels ( B , E ), the blue dots represent the CD11b population that is iNOS − -Arg1 − , the orange dots represent the CD11b population that is iNOS + -Arg1 − and the green dots represent the CD11b population that is double positive for both iNOS + -Arg1 + . These colored dots are also shown in the forward scatter plots of panels ( A , D ).

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Transplantation Assay, Flow Cytometry, Staining, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: CD68 immune cells with a pro-inflammatory Arg1 − iNOS + phenotype after SCI are reduced by SC transplantation. Representative images of flow cytometry analysis and pie charts of CD68 population dynamics at 14 days post-injury (7 days post-transplantation) reveal that CD68 immune cells with a pro-inflammatory Arg1 − iNOS + phenotype after SCI ( A – C ) are reduced by the intraspinal transplantation of SCs ( D – F ). Results are expressed as mean ± SD. For panels ( A , D ), the orange dots represent the ED1 population that were gated based on their forward and side scatter from the total events (blue) that were acquired. The orange dots in the different quadrants of ( B , E ) represent the ED1 population that was positive for either Arg1 (top left quadrant) or iNOS (bottom right quadrant) or double positive for both markers (top right quadrant).

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Transplantation Assay, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: CD11b immune cells expressing a highly pro-inflammatory CD38 + iNOS + phenotype or anti-inflammatory Arg1 + CD163 + form after SCI were unaltered by SC transplantation. Representative images of flow cytometry analysis and pie charts of CD11b population dynamics at 14 days post-injury (7 days post-transplantation) reveal that CD11b immune cells with a highly pro-inflammatory CD38 + iNOS + phenotype after SCI ( A , B ) are not altered by SC transplantation ( E , F ). Similarly, CD11b immune cells with a highly anti-inflammatory Arg1 + CD163 + phenotype were unchanged across SCI controls ( C , D ) and SC-transplanted groups ( G , H ). Results are expressed as mean ± SD. For panels ( A , B ), the blue dots represent the CD11b population that were CD38 − iNOS − , the orange dots represent the CD11b population that were CD38 − iNOS + , the gray dots represent the CD11b population that were CD38 + iNOS − and the green dots represent the CD11b population that were double positive for both CD38 + iNOS + . For panels ( C , D ), the configuration for the colored dots is the same for the representation labeling, single, double or absent, though the proteins Arg1 and CD163 are represented rather than CD38 and iNOS.

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Expressing, Transplantation Assay, Flow Cytometry, Labeling

Journal: International Journal of Molecular Sciences

Article Title: Schwann Cell Transplantation Subdues the Pro-Inflammatory Innate Immune Cell Response after Spinal Cord Injury

doi: 10.3390/ijms19092550

Figure Lengend Snippet: Cytokine profile of CD11b + cells is unaltered by SC implants after SCI. Representative images of flow cytometry analysis and pie charts of CD11b population dynamics at 14 days post-injury (7 days post-transplantation) show that CD11b immune cells have unaltered expression of pro- (TNF-α, IL-1β) and anti-inflammatory (IL-4, IL-10) cytokines comparatively among SCI controls ( A – E ) and SC-transplanted groups ( F – J ). Results are expressed as mean ± SD. For panels ( A – D , F – I ) the blue dots represent the cell population that were CD11b − or did not express the selected cytokines, whereas the green dots represent the CD11b + population that expressed CD11b and the cytokine of interest (top right quadrant).

Article Snippet: Cells were then washed once with flow cytometry staining buffer (R&D System), fixed, and permeabilized (BD Cytofix/Cytoperm, BD Bioscience) for 20 min to permit immunostaining with selected antibodies against intracellular proteins for 1 h on ice.

Techniques: Flow Cytometry, Transplantation Assay, Expressing

Journal: EMBO Reports

Article Title: DJ‐1 depletion prevents immunoaging in T‐cell compartments

doi: 10.15252/embr.202153302

Figure Lengend Snippet: A Representative flow‐cytometry plots of CD44 and CD62L expression on total CD8 T cells of 45‐week‐old Dj‐1 KO and age‐ and sex‐matched WT mice. B, C Percentages of CD44 low CD62L high (Tn) (B) and CD44 high CD62L low (Tem) (C) cells among total CD8 T cells of spleen and pLNs from young and 45‐week‐old Dj‐1 KO and WT littermates (young KO, n = 5; young WT, n = 5; 45‐week‐old KO, n = 8; 45‐week‐old WT, n = 6; for 45‐week‐old mice, data pooled from 2 independent experiments). D Representative histogram overlay of PD‐1 expression among total CD8 T cells in spleen of 45‐week‐old mice (left panel) and percentages of PD‐1 + cells among total CD8 T cells (right panel). E Percentages of Ki‐67 + cells among total CD8 T cells. F Percentage of Ki‐67 + cells among splenic CD8 Tn, Tem, and Tcm in 45‐week‐old Dj‐1 KO and age‐ and sex‐matched WT mice. G Representative flow‐cytometry plots of Ki‐67 and PD‐1 among splenic CD8 Tem of 45‐week‐old mice. H IFN‐γ production in CD8 T cells of spleen and pLNs after in vitro stimulation using 50 ng/ml of PMA and 750 ng/ml of ionomycin for 5 h. I Representative flow‐cytometry plots of CD44 and CD49d among blood CD8 T cells of 60‐week‐old mice. J Percentages of Tn (top), Tvm (middle), and Tmem (bottom) among 60‐week‐old or young mice (young KO, n = 4; young WT, n = 3; 60‐week‐old KO, n = 4; and 60‐week‐old WT, n = 5). K Representative histogram of CD31 among blood CD8 T cells of 60‐week‐old mice. L CD31 MFI of total CD8 T cells (young KO, n = 4; young WT, n = 3; 60‐week‐old KO, n = 4; and 60‐week‐old WT, n = 5). Data information: SP and LN represent spleen and lymph nodes, respectively. Results represent at least four (B‐H) or two (I‐L) independent experiments. Data are mean of biological replicates ± SD. Each dot/symbol represents the measurement from one mouse. The P ‐values are determined by a two‐tailed non‐paired Student’s t ‐test. n.s. or unlabeled, not significant, * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001. Source data are available online for this figure.

Article Snippet: Spleens and peripheral lymph nodes (pLN) were collected and stored in the ice cold flow‐cytometry staining (FCM) buffer [Ca 2+ and Mg 2+ free PBS (Lonza, BE17‐516F) with 2% inactivated fetal bovine serum (FBS) and 2 mM EDTA, pH 8.0].

Techniques: Flow Cytometry, Expressing, In Vitro, Two Tailed Test

Journal: EMBO Reports

Article Title: DJ‐1 depletion prevents immunoaging in T‐cell compartments

doi: 10.15252/embr.202153302

Figure Lengend Snippet: A Expression level of CD31 among total blood or splenocyte CD4 T cells of 45‐week‐old (left) and young (right) mice. B Representative flow‐cytometry plots of CD44 and CD62L expression on total CD4 T cells of 45‐week‐old Dj‐1 KO and age‐ and sex‐matched WT mice (young KO, n = 5; young WT, n = 5; 45‐week‐old KO, n = 8; 45‐week‐old WT, n = 6; for 45‐week‐old mice, data pooled from 2 independent experiments; of note, more than one pLNs might be taken from several mice). C, D Percentages of CD44 low CD62L high (Tn) (C) and CD44 high CD62L low (Tem) (D) cells among total CD4 T cells of spleen and pLNs from young and 45‐week‐old Dj‐1 KO and WT littermates. E Representative histogram overlay of PD‐1 expression among total CD4 T cells in spleen of 45‐week‐old mice (left panel) and percentages of PD‐1 + cells among total CD4 T cells (right panel). F Representative histogram overlay of CTLA‐4 expression among total CD4 T cells in spleen of 45‐week‐old mice (left panel) and percentages of CTLA‐4 + cells among total CD4 T cells (right panel). G Percentages of Ki‐67 + cells among total CD4 T cells. H IFN‐γ production in CD4 T cells of spleen and pLNs after in vitro stimulation using 50 ng/ml of PMA and 750 ng/ml of ionomycin for 5 h. I The selected significantly enriched GO‐terms and pathways among the downregulated genes in CD4 Tconv cells from 45‐week‐old Dj‐1 KO mice versus the age‐ and gender‐matched WT littermates from microarray analysis (upper panel). Lower panel, volcano plot shows both downregulated and upregulated differentially expressed genes in splenic CD4 T cells from three 45‐week‐old Dj‐1 KO mice versus three age‐matched WT littermates. The dashed line in y axis corresponds to the value of 1.3 ( P = 0.05), while the two dashed lines in x ‐axis correspond to −1 and 1 (change fold = 2). A two‐tailed Student t ‐test was used to calculate the P values (for detailed microarray analysis method, refer to Materials and Methods). J, K Comparison of naive CD4 (Tn) mitochondrial mass (mito mass, J) and membrane potential (mito potential, K) of young and 45‐week‐old Dj‐1 KO and WT mice. L, M Comparison of CD4 Tem mitochondrial mass (mito mass, L) and membrane potential (mito potential, M) of young and 45‐week‐old Dj‐1 KO and WT mice. SP and LN represent spleen and lymph nodes, respectively. Data information: results represent at least four (B–G) and three (J–M) independent experiments. Data are mean of biological replicates ± SD. Each biological replicate indicates the measurement from one individual mouse. The P ‐values are determined by a two‐tailed un‐paired Student’s t ‐test. n.s. or unlabeled, not significant, * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001, and **** P ≤ 0.0001.

Article Snippet: Spleens and peripheral lymph nodes (pLN) were collected and stored in the ice cold flow‐cytometry staining (FCM) buffer [Ca 2+ and Mg 2+ free PBS (Lonza, BE17‐516F) with 2% inactivated fetal bovine serum (FBS) and 2 mM EDTA, pH 8.0].

Techniques: Expressing, Flow Cytometry, In Vitro, Microarray, Two Tailed Test

Journal: EMBO Reports

Article Title: DJ‐1 depletion prevents immunoaging in T‐cell compartments

doi: 10.15252/embr.202153302

Figure Lengend Snippet: A Schematic of the experimental setup of bone marrow transplantation. A total of 10E6 of bone marrow cells from young Dj‐1 KO mice (CD45.2 + ) and WT mice (CD45.1 + ) (1:1 mix) were transferred into lethally‐irradiated young WT recipients (CD45.2 + ) by i.v. injection. Mice stably engrafted with donor cells were sacrificed for flow cytometry (FCM) analysis later. B, C Percentages of KLRG1 + CD8 T cells derived from young Dj‐1 KO and WT donor BM cells in blood (B) and spleen (C) within young WT recipients ( n = 5; blood sampled twice at both 6 and 8 weeks). D Percentages of PD‐1 + cells among total CD8 T cells derived from young Dj‐1 KO and WT BM cells in spleen within young WT recipients. E, F Percentages of CD8 Tem derived from young Dj‐1 KO and WT BM cells in blood (E) and spleen (F) within young WT recipients. G, H Ratios between CD8 Tn and Tem cells developed from CD45.1 (WT) or CD45.2 (KO) BM cells in blood (G) and spleen (H) within young WT recipients. I, J Percentages of CD8 Tn in blood (I) and spleen (J) derived from young Dj‐1 KO and WT BM cells within young WT recipients. K, L Percentage of CD8 Tcm among total CD8 T cells derived from young Dj‐1 KO and WT BM cells in blood (K) and spleen (L) of young WT recipients. M Percentages of CD8 single‐positive cells among thymus originated from 45‐week‐old Dj‐1 KO and WT BM cells within young Dj‐1 KO or WT recipients following reconstitution. N, O Percentages of CD8 CD44 high CD62L high (Tcm) cells in blood (N) and spleen (O) derived from 45‐week‐old Dj‐1 KO and WT BM cells within young Dj‐1 KO or WT recipients. Data information: Results represent two independent experiments. The P ‐values are determined by a two‐tailed paired Student’s t ‐test. n.s., not significant, * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001.

Article Snippet: Spleens and peripheral lymph nodes (pLN) were collected and stored in the ice cold flow‐cytometry staining (FCM) buffer [Ca 2+ and Mg 2+ free PBS (Lonza, BE17‐516F) with 2% inactivated fetal bovine serum (FBS) and 2 mM EDTA, pH 8.0].

Techniques: Transplantation Assay, Irradiation, Injection, Stable Transfection, Flow Cytometry, Derivative Assay, Two Tailed Test

Journal: EMBO Reports

Article Title: DJ‐1 depletion prevents immunoaging in T‐cell compartments

doi: 10.15252/embr.202153302

Figure Lengend Snippet: Schematic of the experimental setup of BM transplantation. A total of 10E6 of bone marrow cells from 45‐week‐old Dj‐1 KO mice (CD45.2 + ) and WT mice (CD45.1 + ) (1:1 mix) were transferred into lethally irradiated young Dj‐1 KO or WT recipients (CD45.2 + ) by i.v. injection. Mice stably engrafted with donor cells were sacrificed for flow cytometry (FCM) analysis later. Percentages of KLRG1 + CD8 T cells derived from 45‐week‐old Dj‐1 KO and WT BM cells within young Dj‐1 KO ( n = 4) or WT recipients ( n = 5). Percentages of KLRG1 + among CD8 Tem derived from 45‐week‐old Dj‐1 KO and WT BM cells within young Dj‐1 KO or WT recipients. Percentages of PD‐1 + population among CD8 T cells derived from 45‐week‐old Dj‐1 KO and WT BM cells within young Dj‐1 KO ( n = 4) or WT recipients ( n = 5). Percentages of CD8 + CD44 high CD62L low (Tem) cells derived from 45‐week‐old Dj‐1 KO and WT BM cells within young Dj‐1 KO or WT recipients. Ratios between blood CD8 Tn and Tem cells developed from two types of BM cells within young Dj‐1 KO or WT recipients. Percentages of CD8 + CD44 low CD62L high (Tn) cells derived from 45‐week‐old Dj‐1 KO and WT BM cells within young Dj‐1 KO or WT recipients. Schematic of the experimental setup of CD8 Tn adoptive transfer. 2.5E5 naïve CD8 T cells isolated from young or 55‐week‐old Dj‐1 KO and WT littermates were injected into Rag‐1 deficient mice by i.v. injection. 6 weeks later, mice were sacrificed for FCM (flow cytometry) analysis. Representative flow‐cytometry plots of KLRG1 + population among total CD8 T following adoptive transfer of CD8 Tn from 55‐week‐old mice. Percentages of KLRG1 + population among total CD8 T cells (55‐week‐old KO, n = 4; 55‐week‐old WT, n = 5; young KO, n = 4; and young WT, n = 4). Percentages of KLRG1 + population among CD8 Tem cells. Percentages of KLRG1 + PD‐1 + population among total CD8 T cells (55‐week‐old KO, n = 4; 55‐week‐old WT, n = 5; young KO, n = 4; and young WT, n = 4). Percentages of KLRG1 + PD‐1 + population among CD8 Tem cells. Data information: Results from BM transfer and adoptive transfer of CD8 Tn represent two independent experiments. Data are mean ± SD. The P ‐values are determined by a two‐tailed paired (B–G) or non‐paired (J–M) Student’s t ‐test. n.s. or unlabeled, not significant, * P ≤ 0.05 and ** P ≤ 0.01.

Article Snippet: Spleens and peripheral lymph nodes (pLN) were collected and stored in the ice cold flow‐cytometry staining (FCM) buffer [Ca 2+ and Mg 2+ free PBS (Lonza, BE17‐516F) with 2% inactivated fetal bovine serum (FBS) and 2 mM EDTA, pH 8.0].

Techniques: Transplantation Assay, Irradiation, Injection, Stable Transfection, Flow Cytometry, Derivative Assay, Adoptive Transfer Assay, Isolation, Two Tailed Test

Journal: EMBO Reports

Article Title: DJ‐1 depletion prevents immunoaging in T‐cell compartments

doi: 10.15252/embr.202153302

Figure Lengend Snippet: Representative flow cytometry data of CD69 and Celltrace violet (CTV) staining on gated living CD8 T cells. The purified CD8 Tn were isolated from young mice and stimulated by different doses of anti‐CD3 ab for 72 h. Enlarged number representing the percentage of the corresponding gate. The percentages of proliferating cells among living CD8 singlets (CD8 Tn were from young mice). The percentages of CD69 + cells among living CD8 T singlets (CD8 Tn were from young mice). Representative flow cytometry data of CD69 and CTV staining on gated living CD8 T cells. The purified CD8 Tn were isolated from 45‐week‐old mice and stimulated by different doses of anti‐CD3 ab for 72 h. Enlarged number representing the percentage of the corresponding gate. The percentages of proliferating cells among living CD8 singlets (CD8 Tn were from 45‐week‐old mice). The percentages of CD69 + cells among living CD8 T singlets (CD8 Tn were from 45‐week‐old mice). Data information: Results represent at least two independent experiments (A‐F). Data are mean ± SD. All the CD8 Tn cells were first pooled from 3 to 4 mice of the same group before exposing to different doses of anti‐CD3 ab. The error bar (SD) here essentially refers to technical replicates. The P ‐values are determined by a two‐tailed non‐paired (B‐C, E‐F) Student’s t ‐test. Unlabeled, not significant, * P ≤ 0.05, ** P ≤ 0.01, and *** P ≤ 0.001.

Article Snippet: Spleens and peripheral lymph nodes (pLN) were collected and stored in the ice cold flow‐cytometry staining (FCM) buffer [Ca 2+ and Mg 2+ free PBS (Lonza, BE17‐516F) with 2% inactivated fetal bovine serum (FBS) and 2 mM EDTA, pH 8.0].

Techniques: Flow Cytometry, Staining, Purification, Isolation, Two Tailed Test

Journal: British Journal of Cancer

Article Title: Bone marrow-derived mesenchymal stromal cells promote colorectal cancer cell death under low-dose irradiation

doi: 10.1038/bjc.2017.415

Figure Lengend Snippet: Co-culture system showed attenuate proliferation and viability under UV irradiation. ( A ) This is a representative figure of proliferation experiment. In total, 4 × 10 4 SW1116, HT29 or DanG cells were seeded with or without (control group) 1 × 10 4 MSCs in 12-well plate. In total, 10 J cm −2 irradiation was performed for 1 h and last for 6 h and total cell numbers in each well were counted at 12 h, 36 h, and 72 h respectively. ( B ) CSFE assay of control and co-cultivation group. ( C , D ) 25, 50, 100, or 200 SW1116, SW620 cells were seeded in ultra-low attachment 24-well plates, respectively (control), or following by additional 25 bone marrow-derived mesenchymal stromal cells seeding in each well. Twenty-five more SW1116 cells were also seeded instead of 25 MSCs as blank control. After 10 J cm −2 irradiation 1 h per 8 h for 8 days, cell colonies were counted on the 9th day. Independent experiments were repeated 2∼3 times. Mean value were represented. ( E ) 6 × 10 6 SW1116 cells were seeded in co-culture system , with or without 10 6 MSC cells seeding in the inserted well. 10 J cm −2 irradiation was performed for 1 h and last for 8 h and total cell numbers in each well were counted at 24 h, 48 h, 72 h, 96 h, and 120 h, respectively. The colorectal cancer cells decreased more rapidly in the co-culture group than control group (SW1116 only) at the beginning (0∼96 h) after irradiation, however, turned to be slower and stayed stable after 96 h. ( F ) Co-culture model for MSCs and CRC. The μ –Slide 2 × 9 well harbours two arrays of 3 × 3 square fields where cells can be cultivated independently within the small square or share the same growth medium within the total 3 × 3 square fields. After co-cultivation for 48 h, flow cytometry showed more CD133+CD44+ colorectal cancer stem cell-like cells than the control group. However, there were also more 7-AAD+dead cells compared with the control group. *P<0.05.

Article Snippet: Intracellular staining flow cytometry followed the standard protocol provided by BD.

Techniques: Co-Culture Assay, Irradiation, Control, Derivative Assay, Flow Cytometry

Journal: British Journal of Cancer

Article Title: Bone marrow-derived mesenchymal stromal cells promote colorectal cancer cell death under low-dose irradiation

doi: 10.1038/bjc.2017.415

Figure Lengend Snippet: Alteration of cytokine levels and protein expressions in MSCs or CRC cells from the co-cultivation system. ( A ) Supernatant from MSCs and CRC cells co-cultivation model were collected at 6 h, 12 h, and 24 h after irradiation. ELISA array was performed, R studio was used for calculating and establishing the heat map. ( B – E ) To identify the origin of cytokines, MSCs, and CRCs were collected from the irradiated co-culture system, respectively. Elevated TNFa, IFN-gamma, and CD154 (CD40L) were detected by flow cytometry in MSCs rather than CRC cells. Proportions of cells are shown in column; ( D ): expression of cytokines in MSC from co-cultivated model before and after irradiation; ( E ): cytokines expression of CRC cells in co-cultivated model before and after irradiation; ( F ): PI3K/AKT signal pathway protein p-Akt and p-Erk1/2 from colorectal cancer cells were significantly suppressed in the coculture system after 6 h, 12 h, and 24 h UV irradiation; ( G ): after 10 Gy ionising irradiation and 24 h incubation, colorectal cancer cell line showed significantly increased necrosis rate, in the coculture model tested by 7-AAD/Annexin V cell apoptosis assay. * P <0.05.

Article Snippet: Intracellular staining flow cytometry followed the standard protocol provided by BD.

Techniques: Irradiation, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Flow Cytometry, Expressing, Incubation, Apoptosis Assay

Journal: Neural Regeneration Research

Article Title: Yizhijiannao Granule and a combination of its effective monomers, icariin and Panax notoginseng saponins , inhibit early PC12 cell apoptosis induced by beta-amyloid (25–35) ☆

doi: 10.3969/j.issn.1673-5374.2012.24.001

Figure Lengend Snippet: PC12 apoptosis by flow cytometry. (A) Nerve growth factor group. (B) Model group. (C) Icariin + Panax notoginseng saponins (PNS) group. (D) PNS group. (E) Yizhijiannao Granule-containing serum group. (F) Icariin group. Propidium iodide (PI) is used to label cells in the middle and late stages of apoptosis and Annexin V is used to label early apoptotic cells. Annexin V-fluorescein isothiocyanate (FITC) + /PI cells are early apoptotic cells. The left superior quadrant represents necrotic cells; the left inferior quadrant represents normal cells; the right superior quadrant represents early apoptotic cells; and the inferior quadrant represents non-viable apoptotic cells. Early PC12 apoptosis was significantly reduced in the Yizhijiannao Granule-containing serum and icariin + PNS groups compared with the icariin or PNS groups.

Article Snippet: The early apoptosis rate of PC12 cells was detected using Annexin V-fluorescein isothiocyanate/propidium iodide staining flow cytometry (Becton Dickinson) according to the Annexin V-fluorescein isothiocyanate kit (Bender Med Systems, Vienna, Austria).

Techniques: Flow Cytometry

Journal: Microorganisms

Article Title: SARS-CoV-2-Spike Antibody and T-Cell Responses Elicited by a Homologous Third mRNA COVID-19 Dose in Hemodialysis and Kidney Transplant Recipients

doi: 10.3390/microorganisms10112275

Figure Lengend Snippet: Frequencies of SARS-CoV-2-S- IFN-γ-producing T cells in whole blood as measured by a flow cytometry assay for intracellular cytokine staining in hemodialysis patients ( A ) or kidney transplant recipients ( B ) after receipt of a homologous mRNA COVID-19 third dose (3D) relative to those measured at baseline; p -values for comparisons across groups are shown.

Article Snippet: The enumeration of SARS-CoV-2-S-reactive IFN-γ-producing-CD8 + and CD4 + T cells in fresh whole blood was carried out by flow cytometry for intracellular cytokine staining (BD Fastimmune, Becton Dickinson and Company-Biosciences, San Jose, CA, USA), as previously described [ , ].

Techniques: Flow Cytometry, Staining

Journal: Microorganisms

Article Title: SARS-CoV-2-Spike Antibody and T-Cell Responses Elicited by a Homologous Third mRNA COVID-19 Dose in Hemodialysis and Kidney Transplant Recipients

doi: 10.3390/microorganisms10112275

Figure Lengend Snippet: ( A ) Frequencies of SARS-CoV-2-S- IFN-γ producing T cells in whole blood in hemodialysis patients after receipt of an homologous mRNA COVID-19 third dose (3D), either Comirnaty ® or Spikevax ® , relative to those measured at baseline ( B ). Example of an intracellular cytokine staining (ICS) dot plot showing CD4 and CD8 frequencies at baseline and after receipt of a third dose.

Article Snippet: The enumeration of SARS-CoV-2-S-reactive IFN-γ-producing-CD8 + and CD4 + T cells in fresh whole blood was carried out by flow cytometry for intracellular cytokine staining (BD Fastimmune, Becton Dickinson and Company-Biosciences, San Jose, CA, USA), as previously described [ , ].

Techniques: Staining