Fig 1: Expression of PRDM14 in pancreatitis. (A) Representative IHC images for PRDM14 in acute and chronic pancreatitis in TMA slide. Scale bars in image, 50 µm. (B) Staining scores and P values for PRDM14 in tissues, 13 normal pancreas, 115 PDAC, 36 cancer adjacent, 30 PanIN, six acute pancreatitis, and 58 chronic pancreatitis. Error bars in graphs represent mean ± SD. Student's t-test: **P < 0.01, ns, not significant.
Fig 2: Extensive upregulation of abnormal gametogenesis is found in tumors from p53-/- mice.a Sections of paired normal tissues derived from p53-/- and p53+/+ mice were stained with Oct4 antibody. The relative ratio of obvious Oct4+ cells appeared in the p53-/- mice with different ages was shown. b Sections of paired thymic lymphoma of p53-/- mouse and normal thymus of p53+/+ mouse were stained with H&E or antibodies against the indicated proteins. c Sections of malignant teratoma, spleen, and sarcoma from p53-/- mice were stained with Oct4 antibody. d The pie chart showed the spontaneous tumor types with different percentage in p53-/- mice. The age of tumor formation was counted at the appearance of obvious tumors. The plot showed the age of appearing obvious tumors in the different spontaneous tumor types in p53-/- mice. e The correlation between the appearance of PGC-like cells and tumor formation in a variety of tissues from p53-/- mice. f Section from spontaneous tumors of p53-/- mice were stained with antibody against Prdm14, Nanos3, Vasa, or H&E. Section of sarcoma stained with Vasa antibody showed an oocyte-like tumor cell (arrow). g Sections were stained with H&E (scale bar = 20 µm)
Fig 3: Immunodetection of binding partners of PRDM14. A, Eluents of a Halo pull-down assay were analyzed by Wes using anti-PRDM14, anti-HSP90a, anti-GRP78, anti-?-Catenin, and anti-caspase-14 antibodies. B, C, Co-immunoprecipitation (Co-IP) experiments were carried out with lysates prepared from Halo-PRDM14-transduced HCC1937 and MDA-MB231 cells. The lysates were immunoprecipitated with anti-HSP90a, anti-GRP78, or anti-?-Catenin antibodies and then immunoblotted (IB) with (B) each antibody and (C) anti-Halo antibody. GRP78, glucose-regulated protein 78; HSP, heat shock protein
Fig 4: NanoLuc luciferase-based bioluminescence resonance energy transfer (NanoBRET) analysis in living cells. A, Schematic of NanoBRET assay for detecting intracellular protein-protein interactions by measuring energy transfer between a NanoLuc (NLuc) donor fusion protein and a HaloTag acceptor fusion protein. NLuc with substrate excites fluorescence-conjugated HaloTag ligand with HaloTag fusion protein by energy transfer. B, 293T cells were transfected with Halo-PRDM14 and NLuc fusion proteins. Expression level of Halo-PRDM14 was analyzed using Wes with anti-PRDM14. GAPDH was used as a loading control. Expression levels of NLuc fusion proteins expressed in 293T cells were detected by NLuc substrate. C, D, A simplified NanoBRET donor saturation assay was carried out. 1:1, 1:10, 1:100, and 1:1000 dilutions of NLuc relative to HaloTag DNA were used. HSP90a-NLuc and NLuc-GRP78 were co-transfected with (C) Halo-PRDM14 and (D) Halo-PRDM14-?C measuring energy transfer. Error bars represent the mean ± SD of triplicate samples. GRP78, glucose-regulated protein 78; HSP, heat shock protein
Fig 5: Naïve characterization of NHL-PSCs in transcription level. (a) The heat map shows the expression level of each gene with six primed hiPSC lines and six non-hypoxia leukemia inhibitory factor-dependent pluripotent stem cell (NHL-PSC) lines. Clustering analysis is based on 132 gene expression patterns. GAPDH was used as an internal control and the expression level in each sample was normalized to the expression level in the primed human endometrium (EDOM) cell line. The passage number of NHL-PSCs is indicated as “P”, and the total passage number is indicated. (b) (c) The expression levels of representative naïve and primed cell markers were compared. The average expression levels and standard deviations of each gene in six primed-iPSC lines (grey columns) and six NHL-PSC lines (black columns) are indicated. The data are reported as mean ± SE. Statistically significant differences were identified between primed iPSCs vs. NHL-PSCs using Student's t-test (n = 3). *P < 0.05, **P < 0.01. (d) NHL-EDOM cells expressed the human primed cell markers, PRDM14 and SSEA1. Scale bar is 100 µm.
Supplier Page from Abcam for Anti-PRDM14 antibody