30f11 1 130 116 535 cd41 fitc  (Miltenyi Biotec)

Journal: Advanced Science

Article Title: Spatial Transcriptional Dynamics of CD74⁺ B Cells in Tertiary Lymphoid Structures Drive Immune Evolution in Penile Squamous Cell Carcinoma

doi: 10.1002/advs.202509742

Figure Lengend Snippet: Characterization of B‐cell subtypes from the PSCC spatial transcriptomics data. A) UMAP plot showing the distributions of different B‐cell subtypes. B) Heatmap illustrating the expression levels of markers across B‐cell subtypes. C) Upper panel: spatial distribution of B‐cell subtypes in three representative samples. Lower panel: mIF images of PSCC tissue from the same three representative samples, showing the localization of immune cells (CD45), cancer cells (PanCK), macrophages (CD68), cell membranes, and nuclei (DAPI). Dashed circles indicate TLS regions. D) Proportional representation of B‐cell subpopulations in TLSs compared with all B cells in PSCC tissue. E) Differential gene expression in CD74⁺ B cells, with the highest expression observed for CD74 and HLA‐DRA . The color gradient represents the mean expression levels of the genes. F,G) UMAP plots showing the expression of CD74 and HLA‐DRA in CD74⁺ B cells. H) GO functional enrichment analysis of genes highly expressed in CD74⁺ B cells compared with other subtypes. I) According to the AUCell estimated pathway scores for “GSE12366_GC_BCELL_VS_PLASMA_CELL_UP” (left) and “GSE13411_IGM_MEMORY_BCELL_VS_PLASMA_CELL_UP” (right), CD74⁺ B cells showed transcriptional profiles more closely resembling those of GC B cells and IgM⁺ memory B cells than those of plasma cells. J) Representative IHC images showing the areas of low and high CD74 expression within TLS regions. K) Kaplan–Meier analysis of overall survival in patients with low ( n = 18) versus high ( n = 18) CD74 expression in TLSs, p < 0.05. L) Representative IHC images showing low and high HLA‐DRA expression levels within TLS regions. M) Kaplan–Meier analysis of overall survival in patients with low ( n = 16) versus high ( n = 20) HLA‐DRA expression in TLSs, p < 0.05.

Article Snippet: The antibodies used were CD20 (Proteintech, PE‐FCA65575), CD45 (Proteintech, FITC‐98117), and CD74 (Biolegend, 326812).

Techniques: Expressing, Gene Expression, Functional Assay, Clinical Proteomics

Journal: Nature

Article Title: Fetal hepatocytes protect the HSPC genome via fetuin-A

doi: 10.1038/s41586-024-08307-x

Figure Lengend Snippet: a, b , Representative fluorescence images and positive proportions of γ-H2AX induced by Eto in E12.5 FL LT-HSCs, ST-HSCs and MPPs cultured with or without hepatocytes (refer to Fig. ) ( n = 3). Scale bar, 20 µm. c, d , Representative fluorescence images and positive proportions of γ-H2AX Eto-induced in E12.5 FL-HSPCs in cultures of E16.5 or E12.5 hepatocyte-conditioned medium (co-SHI) or basic medium (SHI) (SHI-control, n = 6, E12.5 Co-SHI, n = 3; E16.5 Co-SHI, n = 5). Scale bar, 20 µm. e , Western-blots analysis of FetuA in SHI and co-SHI media and in E12.5 and E16.5 FL tissues; Coomassie blue was used as a loading control ( n = 3). f , Representative fluorescence images of FetuA in E12.5 FL, E16.5 FL and E19 FL ( n = 3). Scale bar, 500 µm. g , Representative fluorescence images of FetuA and hepatocytes (E-cadherin + ), haematopoietic (CD45/Ter119 + ), endothelial (Scal-1 or CD144 + ), mesenchymal (Nestin + ) cells in E16.5 FL ( n = 3). Scale bars, 10 and 5 µm. h , Representative fluorescence images of FetuA (green) and HSPCs (Lin – CD41 – CD48 – CD150 + ) in E16.5 FL ( n = 3). Scale bars, 10 and 5 µm. i, j , Representative comet-tail DNA images and the proportions of Eto-induced tail DNA in HSPCs cultured with or without FetuA ( n = 3). The medians (red dashed lines) and quartiles (black dashed lines) of each group are shown. Scale bar, 100 µm. k , Representative fluorescence images of FetuA in the E19 bone marrow (BM) of Fetua +/– and Fetua –/– mice. Scale bar, 500 µm. l, m , Representative fluorescence images and positive cell proportions of γ-H2AX (green) in Kit + cells (red) in the E19 BM of Fetua +/– or Fetua –/– mice ( n = 3). Scale bar, 10 µm. n , Representative fluorescence images of FetuA in the E12.5 FL of mice with or without intraplacental (IP) injection of FetuA. Scale bar, 300 µm. o, p , Representative fluorescence images and positive cell proportions of γ-H2AX (green) in Kit + cells (red) in the E12.5 FL of mice with or without IP injection of FetuA followed by Eto-treatment in vivo (IP FetuA–, n = 4; IP FetuA+, n = 3). Scale bar, 10 µm. q , Representative fluorescence images of FetuA in the E16.5 FL of Alb-cre; ROSA26-LSL-DTA mice with or without IP injection of FetuA. Scale bar, 500 µm. r, s , Representative fluorescence images and positive cell proportions of γ-H2AX (green) in Kit + cells (red) in the E16.5 FL of Alb-cre; ROSA26-LSL-DTA mice with or without IP injection of FetuA followed by Eto-treatment in vivo (IP FetuA–, n = 3; IP FetuA+, n = 4). Scale bar, 10 µm. The n represents independent experiments ( a-j ) and fetuses ( k - s ) from 3 independent experiments. The mean ± s.d. is shown ( b, d, m, p, s ). Statistical tests: unpaired two-sided Student’s t-test ( b, d, j, m, p, s ).

Article Snippet: CD45–FITC (104; 1:100, MCD45201) was acquired from Invitrogen.

Techniques: Fluorescence, Cell Culture, Control, Western Blot, Injection, In Vivo