Journal: mSphere

Article Title: Differential innate immune responses of human macrophages and bronchial epithelial cells against Talaromyces marneffei

doi: 10.1128/msphere.00258-22

Figure Lengend Snippet: Cytokine responses of human and murine macrophages cultured in the presence or absence of plasma/serum upon Talaromyces marneffei infection. Upon T. marneffei infection, the relative expression of ( A ) TNF and ( B ) CXCL8 mRNAs in hPBDMs could only be stimulated in the presence of autologous plasma; whereas no significant change in the mRNA expression of these two cytokines was observed when the culture medium was not supplemented with autologous plasma. The infection experiment was conducted in triplicate using blood cells isolated from three independent donors. The inability of T. marneffei to induce ( C ) TNF and ( D ) CXCL8 mRNAs upon infection was also observed in the murine macrophage cell lines J774A.1 and RAW264.7. Data are presented as mean ± SD and comparisons were analyzed using unpaired t -test (two-tailed). **, P < 0.01; ***, P < 0.001.

Article Snippet: Expression of cytokines were measured semi-quantitatively using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instruction; and all samples were tested in duplicates.

Techniques: Cell Culture, Clinical Proteomics, Infection, Expressing, Isolation, Two Tailed Test

Journal: mSphere

Article Title: Differential innate immune responses of human macrophages and bronchial epithelial cells against Talaromyces marneffei

doi: 10.1128/msphere.00258-22

Figure Lengend Snippet: Changes of cytokine mRNA expression in hPBDMs upon Aspergillus fumigatus and Talaromyces marneffei infections. Relative mRNA expressions of CXCL8 and TNF in hPBDMs infected with both fungi were significantly induced mainly after 4 h post-infection and the expression of CXCL10 mRNA in hPBDMs was only significantly induced by T. marneffei infection, in which the expression level reached the peak at 8 h post-infection. The infection experiment was conducted in triplicate using blood cells isolated from three independent donors. The relative mRNA expression levels of CXCL8 , CXCL10 , and TNF were detected by qRT-PCR. Data are presented as mean ± SD and comparisons were analyzed using unpaired t -test (two-tailed). *, P < 0.05; †, P < 0.01; ‡, P < 0.001; ×, undetectable.

Article Snippet: Expression of cytokines were measured semi-quantitatively using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s instruction; and all samples were tested in duplicates.

Techniques: Expressing, Infection, Isolation, Quantitative RT-PCR, Two Tailed Test

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: Caffeic acid selectively eliminates teratogenic human-induced pluripotent stem cells via apoptotic cell death.

doi: 10.1016/j.phymed.2022.154144

Figure Lengend Snippet: Fig. 3. Increase of caspase activities by CAA treatment in iPSCs. (A) Apoptosis-related proteins were detected by Western blotting in CAA-treated iPSCs and hDFs. Relative band intensities were calculated after normalization to β-actin expression. (B) iPSCs and hDFs were treated with 50 and 100 μM CAA for 24 h, and the caspase-3, −8, and −9 activities were determined. Relative caspase activity compared with CAA-untreated iPSCs and hDFs was expressed as the means ± SD (n = 3). (C) iPSCs were pre-treated with or without 10 μM of caspase-3 inhibitor (z-DEVD), caspase-8 inhibitor (z-IETD), caspase-9 inhibitor (z-LEHD), and pan-caspase inhibitor (z-VAD) for 30 min and then treated with 50 and 100 μM CAA. DMSO was treated as vehicle control. After 24 h, cells were stained with crystal violet solution and relative cell viability compared with CAA-untreated control iPSCs were expressed as the means ± SD (n = 3). **p < 0.01 vs. CAA-untreated control. ##p < 0.01 vs. Inhibitor-untreated control. Scale bar = 100 μM.

Article Snippet: The expression profile of human apoptosis-related proteins in CAA-treated or -untreated iPSCs was analyzed using a Proteome Profiler Human Apoptosis Array Kit A. Kim et al. Phytomedicine 102 (2022) 154144 (ARY009, R&D System, Minneapolis, MN, USA) according to the manufacturer’s instruction.

Techniques: Western Blot, Expressing, Activity Assay, Control, Staining

Journal: PloS one

Article Title: Activated human T cells secrete exosomes that participate in IL-2 mediated immune response signaling.

doi: 10.1371/journal.pone.0049723

Figure Lengend Snippet: Figure 3. Cytokine production from autologous exosome stimulated CD3+ T cells at day zero (0 h) and day five (120 h). Relative quantification of spot intensities was performed using Quantity One software (Bio-Rad). Each bar represents an average of the intensity from two protein spots. White bars represent 0 h and grey bars represent 120 h (day 5). The exosomes appeared to contain significant amounts of CCL5 (RANTES) immediately after the addition of exosomes (at 0 h) since the supernatants showed relatively large amounts of RANTES. These levels were decreased at day five. doi:10.1371/journal.pone.0049723.g003

Article Snippet: Analysis of cytokines in the supernatant was carried out using Proteome ProfilerTMArray, Human Cytokine Array Panel A (cat#ARY005, R&D Systems Europe) according to manufacturer’s instructions.

Techniques: Quantitative Proteomics, Software

Journal: PloS one

Article Title: Activated human T cells secrete exosomes that participate in IL-2 mediated immune response signaling.

doi: 10.1371/journal.pone.0049723

Figure Lengend Snippet: Figure 5. Cytokine production from IL-2 stimulated CD3+ T cells at day zero (0 h) and day five (120 h). Relative quantification of spot intensities was performed using Quantity One software (Bio- Rad). Each bar represents an average of the intensity from two protein spots. White bars represent 0 h and grey bars represent 120 h (day 5). Cytokines IL-5, MIF, and GM-CSF (CSF2) were present at a high level in the supernatant after five days. doi:10.1371/journal.pone.0049723.g005

Article Snippet: Analysis of cytokines in the supernatant was carried out using Proteome ProfilerTMArray, Human Cytokine Array Panel A (cat#ARY005, R&D Systems Europe) according to manufacturer’s instructions.

Techniques: Quantitative Proteomics, Software

Journal: PloS one

Article Title: Activated human T cells secrete exosomes that participate in IL-2 mediated immune response signaling.

doi: 10.1371/journal.pone.0049723

Figure Lengend Snippet: Figure 6. Cytokine production from exosome+IL-2 stimulated CD3+ T cells at day zero (0 h) and day five (120 h). Relative quantification of spot intensities was performed using Quantity One software (Bio-Rad). Each bar represents an average of the intensity from two protein spots. White bars represent 0 h and grey bars represent 120 h (day 5). The cytokines IL-5, IL-13 and GM-CSF as well as the chemokines CCL3 and CCL4 were present at higher levels at day five. doi:10.1371/journal.pone.0049723.g006

Article Snippet: Analysis of cytokines in the supernatant was carried out using Proteome ProfilerTMArray, Human Cytokine Array Panel A (cat#ARY005, R&D Systems Europe) according to manufacturer’s instructions.

Techniques: Quantitative Proteomics, Software