Journal: Frontiers in Cell and Developmental Biology

Article Title: Transcriptomic Analysis Identified ARHGAP Family as a Novel Biomarker Associated With Tumor-Promoting Immune Infiltration and Nanomechanical Characteristics in Bladder Cancer

doi: 10.3389/fcell.2021.657219

Figure Lengend Snippet: Knockdown of ARHGAP5-, ARHGAP17-, and ARHGAP24-suppressed BC cell proliferation in vivo and in vitro . (A) System analysis of expression of ARHGAP family gene in BCa cancer cell lines in the CCLE database. (B) ARHGAP5, ARHGAP17, and ARHGAP24 expression in seven pairs of low-grade BCa and seven pairs of high-grade BCa; GAPDH serves as a loading control. (C) ARHGAP5, ARHGAP17, and ARHGAP24 expression in four bladder transitional cell carcinoma cell lines (RT4, UM-UC-3, T24, and 5637) compared with immortalized epithelium cell line (SV-HUC-1). (D,E) Cloning formation of T24, UMUC-3, and ARHGAP5, ARHGAP17, and ARHGAP24 knocking down T24, UMUC-3 (*** p < 0.001 versus shNC). (F,G) 5-Ethynyl-2′-deoxyuridine (EdU) assay of T24 and shARHGAP5-, ARHGAP17-, and ARHGAP24-transfected T24 (*** p < 0.001 versus shNC). (H) Tumor growth size of subcutaneous T24 tumors transfected with shNC or shARHGAP5, ARHGAP17, and ARHGAP24 21 days after tumor implantation. (I) Tumor growth curve of subcutaneous T24 tumors transfected with shNC or shARHGAP5, ARHGAP17, and ARHGAP24 in 21 days of tumor implantation. (J) Average weights of tumors from shNC and shARHGAP5, ARHGAP17, and ARHGAP24 are shown in the histogram. Error bars indicate SD.

Article Snippet: Bladder transitional cell carcinoma cell lines (RT4, UM-UC-3, T24, 5,637, and J82), immortalized uroepithelium cell line (SV-HUC-1), and human embryonic kidney cells (HEK-293) were received from Shanghai Yuanye Bio-Technology Co. (Shanghai, China).

Techniques: Knockdown, In Vivo, In Vitro, Expressing, Control, Cloning, EdU Assay, Transfection, Tumor Implantation

Journal: The Journal of Biological Chemistry

Article Title: Calcium Co-regulates Oxidative Metabolism and ATP Synthase-dependent Respiration in Pancreatic Beta Cells

doi: 10.1074/jbc.M113.513184

Figure Lengend Snippet: Glucose- and calcium-mediated activation of ATP synthase-dependent respiration in human islets. Human islets bound to 804G matrix were incubated in KRBH ( A ) or a KRBH buffer lacking Ca 2+ supplemented with 0.4 m m EGTA ( B ; Ca 2+ -free). For each condition the mean ± S.E. n = 3 from the same donor is shown. Total islet protein varied between wells (4–6 μg). Because of these variations the results are expressed relative to the respiratory rate before glucose stimulation. ATP synthase-dependent ( dep , C ) and -independent respiration ( D ) was quantified as described under “Experimental Procedures.” The results are the mean ± S.E. ( n = 6) obtained from 2 donors. *, p < 0.01; **, p < 0.001; ns , not significant. R , rotenone (1 μ m ); AA , antimycin A (1 μg/ml); O , oligomycin (2.5 μg/ml); glc , glucose. E , preventing calcium signaling blocks glucose-induced insulin secretion. Insulin secretion from human islets was determined in KRBH or the same buffer lacking Ca 2+ in either 1 m m ( gray bars ) or 16.7 m m glucose ( black bars ). Shown is the average ±S.E. n = 4 result with islets from a single donor (*, p < 0.05).

Article Snippet: The 804G bladder carcinoma cell line ( ) and protocols to isolate the matrix were obtained from Viacyte (San Diego, CA).

Techniques: Activation Assay, Incubation