Human MICB Antibody

Detection of Mouse MICB by Flow Cytometry

Hydroxycitrate reduces MICA expression in activated T cells and multiple cancer cells. (A) MICA mRNA analyzed by quantitative RT-PCR in total RNA purified from HEK293 cells after more than 30 passages in glucose (Glc) and galactose (Gal). MICA expression is normalized to housekeeping gene RPLP0 and displayed as mean ± SEM from six independent experiments. (B) MICA surface expression analyzed by flow cytometry of Glc and Gal cells at basal levels. Dot plots are representative of at least three independent experiments. Grid is set to ∼5% of corresponding isotype control stainings. (C) Mitochondrial stress test on HEK293 cultivated in Glc or Gal under same conditions as in Figure 4A. The graph is baselined to measuring point three and displays mean ± SEM from two independent experiments. (D) MICA/B surface expression of peripheral blood lymphocytes (PBLs) activated for 3 days in Glc or Gal growth medium prior to 18 h treatment with FR901228 (20 ng/mL). Grids in dot plots are set to ∼5% of corresponding isotype control staining and dot plots are representative of seven different donors. The bar graph displays mean ± SEM of isotype-corrected MICA/B MFI ( delta MFI) from seven donors. Left panel is zoomed in on the difference between untreated Glc and Gal PBLs. (E,F) HEK293 MGAT5 knockout (KO) cells were treated with (E) 2DG (20 mM) or (F) hydroxycitrate (HC) (15 mM) in addition to PBS (UT), citrate (10 mM), or GlcNAc (25 mM) for 22–24 h. Bar graphs display MICA surface expression as mean ± SEM of delta MFI values from three independent experiments. Data of UT samples share values with UT samples in Figure 3H. (G) MICA surface expression in several cancer cell lines after 18 or 42 h treatment with HC (10 mM). delta MFI values are normalized to UT control and shown as mean ± SEM from at least three independent experiments. (H,I) MICA surface expression (H) and NKG2D-fc binding (I) in cancer cell lines after 2.5 h treatment with HC (10 mM) prior to 18 h stimulation with FR901228 (FR, 20 ng/mL) or sodium butyrate (But, 5 mM). Bar graphs display MICA surface expression as mean ± SEM of delta MFI values (H), or NKG2D-fc surface binding as ± SEM of delta MFI normalized to untreated (UT) control (I), from three independent experiments. Statistical analysis was performed by unpaired t-test with Welch’s correction in (A,E,F), ratio paired t-test in (D), one-sample t-test in (G), and two-way ANOVA with Bonferroni’s multiple comparison test (H,I). *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32849657), licensed under a CC-BY license. Not internally tested by R&D Systems.