Journal: iScience

Article Title: Alternative polarization of resident macrophages improves hyperglycemia-associated male infertility

doi: 10.1016/j.isci.2022.104430

Figure Lengend Snippet: Generation of AAVs that increase JMJD3 specifically in macrophages (A) AAVs carrying an M2-trigger, JMJD3, under a macrophage-specific CD68 promoter was generated. A scramble sequence was used as a control. GFP reporters were also included in the construct for assessment of the transduction efficiency. (B) Cultured RAW264.7 cells successfully transduced with the viruses. GFP channel was shown. (C) RT-qPCR for JMJD3. (D) JMJD3 activity. ∗p < 0.05. N = 5. Scale bars are 50 μm.

Article Snippet: JMJD3 chemiluminescent Assay Kit , BPS Bioscience , #50406.

Techniques: Generated, Sequencing, Construct, Transduction, Cell Culture, Quantitative RT-PCR, Activity Assay

Journal: iScience

Article Title: Alternative polarization of resident macrophages improves hyperglycemia-associated male infertility

doi: 10.1016/j.isci.2022.104430

Figure Lengend Snippet: Orthotopic injection of AAVs in the testis does not alter diabetes (A) Schematic of the study: we assigned male mice into four groups of five each. Group 1: mice only received an intraperitoneal injection of saline (control for STZ). Group 2: mice only received an intraperitoneal injection of STZ. Group 3: mice received an intraperitoneal injection of STZ and an orthotopic injection of AAVs carrying control Scr into the testis one week after STZ. Group 4: mice received an intraperitoneal injection of STZ and an orthotopic injection of AAVs carrying control JMJD3 into the testis one week after STZ. Mice were examined 12 weeks after treatment. (B) Fasting blood glucose. (C) Serum insulin. (D) Beta-cell mass at sacrifice. (E) Representative immunofluorescent images for insulin in mouse pancreas. ∗p < 0.05. NS: non-significant. N = 5. Scale bars are 100 μm.

Article Snippet: JMJD3 chemiluminescent Assay Kit , BPS Bioscience , #50406.

Techniques: Injection

Journal: iScience

Article Title: Alternative polarization of resident macrophages improves hyperglycemia-associated male infertility

doi: 10.1016/j.isci.2022.104430

Figure Lengend Snippet: JMJD3 expression in testicular macrophages improves mating capability of diabetic male mice At the beginning of the 12 weeks after AAV injection (or the beginning of the 13 weeks after STZ injection; or one week before sacrifice), each male C57BL/6 mouse from four groups was caged with four age-matched untreated female C57BL/6 mice for one week before they were removed from the females for final analysis followed by sacrifice. (A) Vaginal mucous pups in females. (B) Pregnancies per male. (C) Litter sizes for successful pregnancies. ∗p < 0.05. NS: non-significant. N = 5.

Article Snippet: JMJD3 chemiluminescent Assay Kit , BPS Bioscience , #50406.

Techniques: Expressing, Injection

Journal: iScience

Article Title: Alternative polarization of resident macrophages improves hyperglycemia-associated male infertility

doi: 10.1016/j.isci.2022.104430

Figure Lengend Snippet: JMJD3 expression in testicular macrophages improves sperm motility Computer-assisted sperm motility analysis was performed. (A) the motile percentage of sperms. (B and C) curvilinear velocity (C) straight line velocity (C). ∗p < 0.05. NS, non-significant. N = 5.

Article Snippet: JMJD3 chemiluminescent Assay Kit , BPS Bioscience , #50406.

Techniques: Expressing

Journal: iScience

Article Title: Alternative polarization of resident macrophages improves hyperglycemia-associated male infertility

doi: 10.1016/j.isci.2022.104430

Figure Lengend Snippet: JMJD3 expression in testicular macrophages increases testosterone (A–D) The levels of LH (A), FSH (B), and testosterone (C) in serum and the levels of testosterone in testis (D). ∗p < 0.05. NS: non-significant. N = 5.

Article Snippet: JMJD3 chemiluminescent Assay Kit , BPS Bioscience , #50406.

Techniques: Expressing

Journal: iScience

Article Title: Alternative polarization of resident macrophages improves hyperglycemia-associated male infertility

doi: 10.1016/j.isci.2022.104430

Figure Lengend Snippet: JMJD3 repolarizes testicular macrophages back to M2-polarized phenotype (A) The digested testicle cells were subjected to CD68 and GFP double FACS, showing that GFP was exclusively detected in CD68 + cell fraction. (B) RT-qPCR for GFP in testis, blood, liver, lung, and kidney in mice. (C and D) Testicular macrophages were isolated and subjected to flow cytometry. (C) Quantification for % CD163 + cells in total CD68 + cells. (D) Representative flow charts for CD68 and CD163. (E) RT-qPCR for M2 markers (CD206 and Arginase 1 (ARG1)) and M1 markers (iNOS, IL-6, and TNFα) in testicular CD68 + macrophages. ∗p < 0.05. NS, non-significant. N = 5.

Article Snippet: JMJD3 chemiluminescent Assay Kit , BPS Bioscience , #50406.

Techniques: Quantitative RT-PCR, Isolation, Flow Cytometry

Journal: iScience

Article Title: Alternative polarization of resident macrophages improves hyperglycemia-associated male infertility

doi: 10.1016/j.isci.2022.104430

Figure Lengend Snippet:

Article Snippet: JMJD3 chemiluminescent Assay Kit , BPS Bioscience , #50406.

Techniques: Recombinant, Plasmid Preparation, Software

Journal: Nature Communications

Article Title: BMP4 drives primed to naïve transition through PGC-like state

doi: 10.1038/s41467-022-30325-4

Figure Lengend Snippet: a Flow chart for the scRNA-seq analysis of BMP4 induced Primed-Naïve Transition (BiPNT). scRNA-seq experiment was performed with 10x Genomics. b t-SNE projection of all 50406 individual cells during the whole BiPNT process. c Expression of genes in 2 different categories at the indicated time points. d t-SNE projection of cells and typical gene expressions at stage 1. e t-SNE projection of cells and typical gene expressions at stage 2. f Percentages of naïve cells and trophoblast-like cells at different time points. g Representative images of ΔPE- Oct4 -GFP/ Gata2 -tdTomato EpiSC induced BiPNT at D6 and D8. Scale bars, 100 μm. The experiments were repeated independently three times with similar results.

Article Snippet: Fig. 1 Single-cell analysis for BiPNT. a Flow chart for the scRNA-seq analysis of BMP4 induced Primed-Naïve Transition (BiPNT). scRNA-seq experiment was performed with 10x Genomics. b t-SNE projection of all 50406 individual cells during the whole BiPNT process. c Expression of genes in 2 different categories at the indicated time points. d t-SNE projection of cells and typical gene expressions at stage 1. e t-SNE projection of cells and typical gene expressions at stage 2. f Percentages of naïve cells and trophoblast-like cells at different time points. g Representative images of ΔPE- Oct4 -GFP/ Gata2 -tdTomato EpiSC induced BiPNT at D6 and D8.

Techniques: Expressing