Journal: Molecular Systems Biology

Article Title: Molecular profiling reveals features of clinical immunity and immunosuppression in asymptomatic P. falciparum malaria

doi: 10.15252/msb.202110824

Figure Lengend Snippet: PBMCs from P. falciparum symptomatic ( n = 16) and asymptomatic ( n = 24) infected individuals as well as healthy immune controls ( n = 24) were stained with a panel of metal‐labeled antibodies and analyzed by CyTOF. Manual gating was used to select the following populations before FlowSOM clustering: A Individual T H1 ‐like memory CD4 + T cells (CD19 − CD3 + CD4 + CD45RA − CCR6 − CXCR3 + ), T H2 ‐like memory CD4 + T cells (CD19 − CD3 + CD4 + CD45RA − CCR6 − CXCR3 − ), circulating memory T FH cells (CD19 − CD3 + CD4 + CD45RA − CXCR5 + ) memory CD4 + T cells. B Classical (CD3 − CD19 + CD20 + CD10 − CD27 + CD21 + ), atypical (CD3 − CD19 + CD20 + CD10 − CD27 − CD21 − ), and activated MBCs (CD3 − CD19 + CD20 + CD10 − CD27 + CD21 − ).

Article Snippet: 158Gd‐conjugated mouse anti‐human CD10 , Fluidigm , Clone HI10a; Cat# 3158011B.

Techniques: Infection, Staining, Labeling

Journal: Molecular Systems Biology

Article Title: Molecular profiling reveals features of clinical immunity and immunosuppression in asymptomatic P. falciparum malaria

doi: 10.15252/msb.202110824

Figure Lengend Snippet: PBMCs from P. falciparum symptomatic ( n = 16) and asymptomatic ( n = 24) infected individuals as well as healthy immune controls ( n = 24) were stained with a panel of metal‐labelled antibodies and analyzed by CyTOF. t‐distributed Stochastic Neighbor Embedding (tSNE) analysis was performed and FlowSOM clustering was used to identify individual cell subpopulations within gated: T H1 ‐like memory CD4 + T cells (CD19 − CD3 + CD4 + CD45RA − CCR6 − CXCR3 + ) T H2 ‐like memory CD4 + T cells (CD19 − CD3 + CD4 + CD45RA − CCR6 − CXCR3 − ) Circulating memory T FH cells (CD19 − CD3 + CD4 + CD45RA − CXCR5 + ) Classical MBCs (CD3 − CD19 + CD20 + CD10 − CD27 + CD21 + ) Atypical MBCs (CD3 − CD19 + CD20 + CD10 − CD27 − CD21 − ) Activated MBCs (CD3 − CD19 + CD20 + CD10 − CD27 + CD21 − ) The tSNE plots in the top panel display cell density and represent the pooled data for each group, while the lower panel shows a projection of the FlowSOM clusters on a tSNE plot. Heatmaps show the median marker expression for each FlowSOM cluster.

Article Snippet: 158Gd‐conjugated mouse anti‐human CD10 , Fluidigm , Clone HI10a; Cat# 3158011B.

Techniques: Infection, Staining, Marker, Expressing

Journal: Molecular Systems Biology

Article Title: Molecular profiling reveals features of clinical immunity and immunosuppression in asymptomatic P. falciparum malaria

doi: 10.15252/msb.202110824

Figure Lengend Snippet:

Article Snippet: 158Gd‐conjugated mouse anti‐human CD10 , Fluidigm , Clone HI10a; Cat# 3158011B.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Marker, Infection, Purification, Flow Cytometry, Blocking Assay, Generated, Recombinant, Saline, Staining, Software, Cytometry

Journal: MedComm

Article Title: Novel aspect of neprilysin in kidney fibrosis via ACSL4‐mediated ferroptosis of tubular epithelial cells

doi: 10.1002/mco2.330

Figure Lengend Snippet: Genetic knockdown of neprilysin improved the fibrotic and ferroptotic phenotype in TGFβ1‐induced TCMK‐1 cells. (A and B) The knockdown efficiency of NEP siRNA (siNEP) in TCMK‐1 cells evaluated by RT‐PCR and Western blot analysis. (C) Representative Western blot images and quantitative analysis of NEP expression in TCMK‐1 cells. (D) RT‐PCR was performed to detect the mRNA level of Fn, α‐SMA, and Col Iα1. (E) RT‐PCR was performed to detect the mRNA level of ACSL4, GPX4, and Ptgs2 (COX2). (F) Western blot analysis was performed to detect protein expression of Fn, α‐SMA, and Col Iα1. (G) Western blot analysis was performed to detect protein expression of ACSL4, GPX4, and COX2. (H) The accumulation of lipid peroxidation in TCMK‐1 cells was analyzed by BODIPY C11 staining (×200, scale bar = 50 μm). NS p > 0.05, versus control. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, versus siNC. ### p < 0.001, ## p < 0.01, # p < 0.05, versus siNC+TGF‐β1. siNC, negative control.

Article Snippet: Primary antibodies against NEP (1:200, 18008‐1‐AP; Proteintech, USA), α‐SMA (1:100, ET1607‐43; Huabio, China), and ACSL4 (1:200, 81196‐1‐RR; Proteintech) were used in the study.

Techniques: Knockdown, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Staining, Control, Negative Control

Journal: MedComm

Article Title: Novel aspect of neprilysin in kidney fibrosis via ACSL4‐mediated ferroptosis of tubular epithelial cells

doi: 10.1002/mco2.330

Figure Lengend Snippet: Neprilysin aggravated fibrosis by ACSL4‐mediated ferroptosis in TGF‐β1‐induced TCMK‐1 cells. (A and B) The efficiency of NEP overexpression in TCMK‐1 cells evaluated by RT‐PCR and Western blot analysis. (C and D) The overexpression efficiency of ACSL4 in TCMK‐1 cells evaluated by RT‐PCR and Western blot analysis. (E and F) The knockdown efficiency of ACSL4 siRNA in TCMK‐1 cells evaluated by RT‐PCR and Western blot analysis. (G) RT‐PCR was performed to detect the mRNA expression of ACSL4, GPX4, and Ptgs2 (COX2). (H) Western blot analysis was performed to detect protein expression of ACSL4, GPX4, and COX2. (J) RT‐PCR was performed to detect the mRNA expression of Fn, α‐SMA, and Col Iα1. (I) Western blot analysis was performed to detect protein expression of Fn, α‐SMA, and Col Iα1. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05, versus oeNC or siNC or oeNC+siNC. #### p < 0.0001, ### p < 0.001, ## p < 0.01, # p < 0.05, versus oeNC+siNC+TGF‐β1. $$$$ p < 0.0001, $$ p < 0.01, $ p < 0.05, versus oeNEP+siNC+TGF‐β1. siNC, silencing negative control; siNEP, NEP siRNA; siACSL4, ACSL4 siRNA; oeNC, overexpression negative control; oeNEP, overexpression of NEP; oeACSL4, overexpression of ACSL4.

Article Snippet: Primary antibodies against NEP (1:200, 18008‐1‐AP; Proteintech, USA), α‐SMA (1:100, ET1607‐43; Huabio, China), and ACSL4 (1:200, 81196‐1‐RR; Proteintech) were used in the study.

Techniques: Over Expression, Reverse Transcription Polymerase Chain Reaction, Western Blot, Knockdown, Expressing, Negative Control

Journal: Annals of Translational Medicine

Article Title: Cinacalcet attenuated bone loss via inhibiting parathyroid hormone-induced endothelial-to-adipocyte transition in chronic kidney disease rats

doi: 10.21037/atm.2019.06.44

Figure Lengend Snippet: CINA treatment inhibits the EndMT in CKD rats. (A,B) Representative western blot analysis and quantification of levels of the CD31, FSP1, α-SMA, CD10, and CD44 proteins in the different groups of rats, as indicated. The data are presented as the mean ± SD. n=4 for each group. *, P<0.05 vs. the CTL group; #, P<0.05 vs. the CKD group. (C) Representative images of immunofluorescence staining for CD31, FSP1, and CD44 in the bone marrow of rats (upper panel: green for CD31, red for FSP1; lower panel: red for CD31, green for CD44). White arrows indicate colocalization between CD31 and either FSP1 or CD44. DAPI (blue) labels the nuclei. Scale bar, 50 µm. CTL, control; CKD, chronic kidney disease; CINA, cinacalcet; SD, standard deviation.

Article Snippet: PVDF membranes were incubated overnight with primary antibodies against CD31 (sc-376764, Santa Cruz, 1:1,000 dilution), FSP1 (ab-197896, Abcam, dilution), α-SMA (ab-7817, Abcam, 1:1,000 dilution), CD10 (NBP2-15771, Novus Biologicals, 1:1,000 dilution), CD44 (ab-157107, Abcam, 1:1,000 dilution), C/EBP-α (sc-365318, Santa Cruz, 1:1,000 dilution), or PPAR-γ (ab-209350, Abcam, 1:1,000 dilution) followed by horseradish peroxidase-labeled secondary IgG (Santa Cruz).

Techniques: Western Blot, Immunofluorescence, Staining, Control, Standard Deviation

Journal: Frontiers in Medicine

Article Title: PD-1 combined with TRBC1 and pan-T cell antibodies for robustly monitoring angioimmunoblastic T-cell lymphoma

doi: 10.3389/fmed.2022.962428

Figure Lengend Snippet: Flow cytometric findings in non-T cell lymphoma. (A) CD4+ T cells with CD10 and bright PD-1 expression were observed in follicular lymphoma. (B) CD4+ T cells with CD200 and bright PD-1 expression were seen in tuberculous granuloma. (C) Expansion of CD4+T cells with bright PD-1 expression was present in nodular lymphocyte-predominant Hodgkin lymphoma. (D) CD4+ T cells with distinctive bright PD-1 expression were observed in reactive follicular hyperplasia. TRBC1 was polyclonal in these populations.

Article Snippet: PD-1 (EH12.2H7, BioLegend, APC), CD10, TRBC1 (JOVI.1, Caprico biotechnologies, FITC) and some pan-T cell antibodies (such as CD3, CD4, CD5, CD7, and CD8) were placed in an additional tube to confirm or exclude a supposed PTCL diagnosis and for patients with an equivocal clinical history or immunophenotyping.

Techniques: Expressing

Journal: Frontiers in Medicine

Article Title: PD-1 combined with TRBC1 and pan-T cell antibodies for robustly monitoring angioimmunoblastic T-cell lymphoma

doi: 10.3389/fmed.2022.962428

Figure Lengend Snippet: Some AITL immunophenotyping could not be detected by the T-cell screen tube. (A,D,E,F) show BM specimens. (B,C) show tissue samples. (A) Aberrant T cells were CD3+, CD4+, and CD10+ with bright PD-1 expression, and CD7 was partially expressed. (B) Neoplastic T cells were CD4+, CD7+, and CD10+ with slightly diminished CD3 and CD5 expression and bright PD-1 expression. (C) Tumor cells were CD3+, CD5+, CD7+, and CD200+ with diminished CD4 expression and bright CD2, CD56, and PD-1 expression, and CD10 was negative (not shown). (D) Abnormal T cells were CD3+, CD4+, CD5+, and CD200+ with bright CD7 and PD-1 expression, and CD10 was negative. (E) An extremely low ratio of CD7 negative T cells was CD3+, CD4+, and CD10+ with bright CD5 and PD-1 expression. The immunophenotyping of neoplastic T cells shifted over time (E,F) before and after therapy, respectively. TRBC1 did not show polyclonal expression in these populations.

Article Snippet: PD-1 (EH12.2H7, BioLegend, APC), CD10, TRBC1 (JOVI.1, Caprico biotechnologies, FITC) and some pan-T cell antibodies (such as CD3, CD4, CD5, CD7, and CD8) were placed in an additional tube to confirm or exclude a supposed PTCL diagnosis and for patients with an equivocal clinical history or immunophenotyping.

Techniques: Expressing