Fig. 6: Inhibition of CER and S1P synthesis alters the M1- and M2-driven effects on post-hepatectomy hepatocytes, respectively. | Cell Death & Disease

Fig. 6: Inhibition of CER and S1P synthesis alters the M1- and M2-driven effects on post-hepatectomy hepatocytes, respectively.

From: Ceramides and sphingosine-1-phosphate mediate the distinct effects of M1/M2-macrophage infusion on liver recovery after hepatectomy

Fig. 6

M1 BMDMs and M2 BMDMs were transfected with siRNA-Cers2 (siCers2) and siRNA-Sphk1 (siSphk1), respectively. A scramble siRNA was used as a control siRNA (siCON). Primary hepatocytes were isolated from the remnant liver at POD2 and incubated with CM from the transfected BMDM, then the proliferation and apoptosis of hepatocytes were examined. A, B Transfection was performed at 24 h before induction of polarization. mRNA (A) and protein (B) levels of Cers2 and Sphk1 were determined at 48 h after polarization. C–F BMDMs with siRNA transfection and their medium were collected to measure the levels of CER metabolites, including d18:1-CER (C and E), d18:1-SPH, and d18:1-S1P (D and F). G Hepatocytes were fixed and stained with proliferation marker PCNA and apoptosis maker C-Caspase 3. Top: representative images of immunofluorescence staining for PCNA (green). Bottom: representative images of immunofluorescence staining for C-Caspase 3 (green). Nuclei were counterstained with DAPI (blue). H, I PCNA-positive hepatocytes and C-Caspase 3-positive hepatocytes were enumerated to evaluate the proliferation (H) and apoptosis (I) in hepatocytes. J The viability of hepatocytes was measured by MTT assay. Images in B and G represent results from three independent experiments. Data in A, C–F, and H–J were demonstrated as mean ± SD, n = 3; *p < 0.05, **p < 0.01, ***p < 0.001.

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