Journal: iScience

Article Title: Long trimer-immunization interval and appropriate adjuvant reduce immune responses to the soluble HIV-1-envelope trimer base

doi: 10.1016/j.isci.2024.108877

Figure Lengend Snippet:

Article Snippet: CXCL13 levels in NHP plasma were measured using Human CXCL13/BLC/BCA-1 Quantikine ELISA Kit (bio-techne/R&D systems) following instruction, with Human CXCL13 protein at 2-fold dilutions start at 500pg/ml as the standard for quantification.

Techniques: Virus, Neutralization, Recombinant, Transfection, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software

Journal: JCI Insight

Article Title: Spatial transcriptomics identifies candidate stromal drivers of benign prostatic hyperplasia

doi: 10.1172/jci.insight.176479

Figure Lengend Snippet: ( A – D ) RISH exemplified for a laser-microdissected hub, Hub-1. ( E – K ) RISH exemplified for an independent hub, Hub-7. ( A and E ) H&E stain. Inner stroma (yellow star) and outer stroma (blue star) are indicated; scale bar is 500 μm, and all figure insets are an additional 2.7× magnification. ( B , F , and I ) IGF1 expression by RISH (brown staining). ( C , G , and J ) CXCL13 expression by RISH (brown staining). ( D , H , and K ) C7 expression (marks interstitial prostate fibroblasts) by RISH (brown staining); arrows indicate fibroblasts interspersed within the “outside” stroma. Note, RISH for IGF1 , CXCL13 , and C7 was conducted on 7 additional BPH hubs .

Article Snippet: BMS-754807 and PPP were obtained from SelleckChem and human recombinant CXCL13 from PeproTech.

Techniques: Staining, Expressing

Journal: JCI Insight

Article Title: Spatial transcriptomics identifies candidate stromal drivers of benign prostatic hyperplasia

doi: 10.1172/jci.insight.176479

Figure Lengend Snippet: ( A – D ) Single-cell RNA-Seq of paired BPH-normal human prostate tissue. ( A ) t-Distributed stochastic neighbor embedding (t-SNE) plot of combined BPH/normal prostate cells. Each dot represents an individual cell. Cell clusters (colored) are annotated by the expression of known cell type markers (e.g., LUM and DCN in fibroblasts). ( B ) Close-up of fibroblast cells contributed by BPH (purple) or normal prostate (green). ( C ) Fibroblast gene expression levels shown for IGF1 , CXCL13 , SRD5A2 , and AR . Color bar depicts log 2 transcript counts per cell. ( D ) Cell coexpression (purple) of IGF1 with CXCL13 , SRD5A2 , or AR . ( E – J ) Two-color RNA in situ hybridization (RISH) of BPH tissue, exemplified for Hub-8, verifies cell coexpression of ( E and H ) IGF1 (blue) with CXCL13 (red); ( F and I ) IGF1 (blue) with SRD5A2 (red); and ( G and J ) IGF1 (blue) with AR (red). Arrows identify representative cells with dual staining; scale bar is 500 μm, and all figure insets are an additional 8.7× magnification. Note, 2-color RISH for IGF1 / CXCL13 , IGF1 / SRD5A2 , and IGF1 / AR was conducted on 1 additional BPH hub .

Article Snippet: BMS-754807 and PPP were obtained from SelleckChem and human recombinant CXCL13 from PeproTech.

Techniques: RNA Sequencing Assay, Expressing, RNA In Situ Hybridization, Staining

Journal: JCI Insight

Article Title: Spatial transcriptomics identifies candidate stromal drivers of benign prostatic hyperplasia

doi: 10.1172/jci.insight.176479

Figure Lengend Snippet: ( A ) H&E stain of Hub-8. Inner stroma (yellow star) and outer stroma (blue star) are indicated; scale bar is 500 μm. ( B – F ) Two-color RNA in situ hybridization (RISH) of Hub-8 shown for ( B and E ) IGF1 (blue) and IGF1R (red), ( C and F ) CXCL13 (blue) and CXCR5 (red), and ( D ) negative control probes. Note, 2-color RISH for IGF1 / IGF1R and CXCL13 / CXCR5 was conducted on 3 additional BPH hubs . ( G ) IHC of Ki-67 identifies proliferating ductal epithelial cells adjacent to IGF1 + stroma. Arrows identify representative Ki-67 + cells. ( H ) Increased proliferating (Ki-67 + ) ductal epithelial cells on the side facing the inner/inductive stroma. Data from 4 hubs (see , quantified as Ki-67 + cells per millimeter duct; scale bar is 500 µm, and all figure insets are an additional 1.6× magnification. Mean ± SD shown. * P < 0.05; 2-sided paired Student’s t test. ( I ) Two-color IHC identifies B cells (CD20, brown) and T cells (CD3, red) within both the inner and outer stroma of hubs. Note, 2-color IHC for CD20/CD3 was conducted on 1 additional BPH hub (Hub-7).

Article Snippet: BMS-754807 and PPP were obtained from SelleckChem and human recombinant CXCL13 from PeproTech.

Techniques: Staining, RNA In Situ Hybridization, Negative Control

Journal: JCI Insight

Article Title: Spatial transcriptomics identifies candidate stromal drivers of benign prostatic hyperplasia

doi: 10.1172/jci.insight.176479

Figure Lengend Snippet: Shown are cross sections of human fetal prostates at gestational ages ( A – C ) 12 weeks showing the urethra, ( D – F ) 14.6 weeks showing ductal budding from the urethra, and ( G – I ) 15 weeks showing ductal branching. ( A , D , and G ) H&E stains; scale bar is 500 μm. Note, the large black circle in G is a bubble under the coverslip. ( B , E , and H ) Two-color RNA in situ hybridization (RISH) of IGF1 (blue) and IGF1R (red). ( C , F , and I ) Two-color RISH of CXCL13 (blue) and CXCR5 (red). Arrows mark occasional epithelial cells with low-intensity CXCL13 and/or CXCR5 expression; scale bar is 500 µm, and all figure insets are an additional 3.2×–4.5× magnification.

Article Snippet: BMS-754807 and PPP were obtained from SelleckChem and human recombinant CXCL13 from PeproTech.

Techniques: RNA In Situ Hybridization, Expressing

Journal: JCI Insight

Article Title: Spatial transcriptomics identifies candidate stromal drivers of benign prostatic hyperplasia

doi: 10.1172/jci.insight.176479

Figure Lengend Snippet: ( A – E ) BPH-1 cell spheroids grown in Matrigel to 10 days and ( F – J ) patient-derived BPH organoids grown in Matrigel to 14 days, in media containing ( A and F ) vehicle (DMSO), ( B and G ) IGF1R inhibitor BMS-754807 (1 μM), ( C and H ) IGF1R inhibitor picropodophyllin (PPP) (1 μM), or ( D and I ) recombinant human CXCL13 (100 ng/mL). Scale bar is 100 μm. ( E ) BPH-1 spheroid and ( J ) BPH organoid generation quantified as the largest 25 diameters (μM) in a random 10× microscope field. Mean ± SD shown. *** P < 0.001; 2-sided Student’s t test, with Bonferroni’s correction applied for multiple comparisons. Each experiment was conducted twice. “Ctr” is media-alone comparison control for CXCL13.

Article Snippet: BMS-754807 and PPP were obtained from SelleckChem and human recombinant CXCL13 from PeproTech.

Techniques: Derivative Assay, Recombinant, Microscopy, Comparison