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Figure 4. CHME-5 cells are CD68-positive and GFAP-negative. CHME-5 cells were stimulated with LPS (1 μg/mL) at 37 °C for 3, 6, and 18 h. A: CD68 mRNA expression was analyzed using RT-PCR analysis with β-actin as housekeeping gene, 3 h (****P < 0.0001) and 6 h (**P < 0.01) vs. US. Image is representative of 3 independent experiments (n = 3) for each treatment group; B: CHME-5 cells were stimulated with LPS (1 μg/mL) at 37 °C for 10 min, whole cell lysates were subjected to SDS-PAGE electrophoresis and immunoblotted with CD68 (1:500) and β-tubulin (1:1,000) antibodies; C: integrated density, ***P < 0.002 vs. US. Image is representative of 4 independent experiments (n = 4) for each group; D: CHME-5 and NHA cells were stimulated with LPS (1 μg/mL) at 37 °C for 10 min, whole cell lysates were subjected to SDS-PAGE electrophoresis and immunoblotted with GFAP (1:2,000) and β-tubulin (1:1,000) antibodies; E: integrated density, (***P < 0.002, ****P < 0.0001) vs. CHME-5. Image is representative of 3 independent experiments (n = 3) for each treatment group. Bars are presented as mean ± SEM. CD68: cluster of differentiation 68; GFAP: glial fibrillary astrocytic protein; LPS: lipopolysaccharide; RT-PCR: real-time polymerase chain reaction; US: unstimulated; SDS-PAGE: sodium dodecyl sulfate polyacrylamide gel electrophoresis