mRNA expression of LSSIG and GPR43.

(A-H) Northern blot analysis. The membrane was hybridized with the probes of LSSIG (A-E) and GPR43 (F-H). M1 (A) and M1/Y705F (B) cells were stimulated with LIF for the indicated time. (C) M1/STAT3ER cells were stimulated with 4HT for 3 hours. (D) Several cell lines were stimulated for 3 hours with IL-6, interferon γ (IFNγ), LPS, IFNγ plus LPS, LIF, or IL-3. DA1.a and 32D cells were starved for cytokines for 5 hours and were stimulated. C: M1 cells stimulated with LIF for 24 hours as a control. (E) Murine hematopoietic tissues were stimulated for 3 hours. Bone marrow cells (BM), spleen cells (SP), and peritoneal macrophages (PM) were stimulated with IL-6 or LPS. C: M1 cells stimulated with LIF for 24 hours as a control. (F) HL-60 and U937 cells were stimulated with PMA for the indicated time. (G) Human tissue blot. SP, spleen; LN, lymph nodes; TH, thymus; PL, peripheral blood leukocytes; BM, bone marrow; FL, fetal liver. In each lane, 2 μg polyA+RNA was blotted. (H) Human monocytes and neutrophils were stimulated for 3 hours with GM-CSF (GM), IL-4, LPS, or IL-8. C: U937 cells were stimulated with PMA for 6 hours as a control. (The transcripts are indicated by arrowheads. EB indicates ethidium bromide staining of gel; G3PDH, the same membrane reblotted with the probe of G3PDH. Similar results were obtained in more than 3 independent experiments.) (I) Quantitative RT-PCR analysis of GPR43 expression in U937 cells (i) and human monocytes (ii). (i) 1, nonstimulated; 2, stimulated with PMA for 24 hours. (ii) 1, nonstimulated; 2, stimulated with GM-CSF for 3 hours; 3, stimulated with LPS for 3 hours. Values were corrected by the data of RT-PCR for glyceraldehyde phosphate dehydrogenase and represent means of 3 samples. The error bars represent the standard deviations.