Fig. 5: Promotion of B-cell lymphoma tumorigenesis by P4HA2 in stromal fibroblasts.

A–D Injection of luciferased Eµ-myc arf^−/− B cells into C57BL/6 WT and P4ha2^−/− mice. Bioluminescence imaging (A, C) of mice for 18 days after injection. Luminescence intensity curves (B, D) for mice injected with luciferased Eµ-myc arf^−/− B cells. E P4HA2 is located in stromal fibroblasts. Immunofluorescence staining of α-SMA, P4HA2 and DAPI in C57BL/6 WT mice xenograft. Scale bars: 100 μm. F Primary stromal fibroblasts were isolated from bone marrow of C57BL/6 WT and P4ha2^−/− mice. qRT-PCR analysis of Gli2, Ptch1, Ptch2 and HH ligands transcripts was performed using RNA isolated from the primary stromal fibroblasts. G Differential genes of Hh downstream secreted factors obtained from RNA-seq analysis of primary bone marrow stromal fibroblasts. Data are shown as the mean ± SEM (n = 3). *P < 0.1, **P < 0.01, ***P < 0.001. H The growth curve of Eµ-myc arf^−/− B cells co-cultured with primary bone marrow stromal fibroblasts supernatant. WT and P4ha2^−/− primary bone marrow fibroblasts were isolated and cultured. After cell counting, both primary cells were cultured at the same density, and the cultured supernatant was collected for co-culture with Eµ-myc arf^−/− B cells. The cell viability was detected every 12 hr. I qRT-PCR analysis of Gli1 and Gli2 transcripts was performed using the tissue of C57BL/6 WT and P4ha2^−/− mice xenograft. Data are shown as the mean ± SEM (n = 3). ***P < 0.001, ****P < 0.0001. All experiments were repeated three times independently.