Intestinal Ces1/Es-x expression is regulated by nutritional status.

<p>(A) Intestinal Ces1/Es-x protein expression in different nutritional states. Mice were fasted for 24 h and refed for 6 h. Fasted and refed mice were euthanized at 8:00 P.M. TGH is a related carboxylesterase migrating at a lower M_r due to lesser glycosylation and is recognized by the polyclonal anti-Ces1/Es-x antibodies. Two µg intestinal proteins were subjected to analysis. Cnx, calnexin  =  loading control. (B) Quantitation of Ces1/Es-x immunoreactive bands obtained in different nutritional states. **p<0.01 Refed vs Fasted. (C) Ces1/Es-x protein distribution along the small intestine. Small intestine was cut into 12 pieces 2-cm long. Proteins were separated by SDS-PAGE and immunodetected with anti-Ces1/Es-x antibodies. Representative data from 3 different independent experiments are shown. (D) Absence of Ces1/Es-x protein (immunoblot) in the intestine from <i>Ces1/Es-x</i>^−/− mice. (E) H&E staining of small intestine (200×) sections.</p

Decreased intestinal TG accumulation and absence of fat malabsorption in <i>Ces1/Es-x^−/−</i> mice.

<p>(A) Uptake of dietary fat along the length of the small intestine. Mice were fasted for 4 h and gavaged with radiolabeled triolein diluted in olive oil. Two hours later, the small intestine was excised, flushed, cut, digested and radioactivity associated with the intestinal segments was determined. N = 3 mice/group. (B) Fat in stools. Sudan III staining of stool fat upon spontaneous defecation in fed mice. (C) Analysis of fat in stools by organic extraction after spontaneous defecation. N = 5–7 mice/group. (D) Fecal lipids analyzed by TLC. N = 5–7 mice/group.</p

Chylomicrons from <i>Ces1/Es-x^−/−</i> mice present with abnormal composition.

<p>(A) Immunoblotting showing plasma apolipoprotein B composition from mice injected with P-407. Plasma samples were prepared at the indicated times. Representative data from 3 independent experiments. (B) Immunoblotting showing apolipoprotein composition of chylomicrons isolated from wild-type and <i>Ces1/Es-x^−/−</i> mice. Representative data from 3 independent experiments. (C) Chylomicron lipid composition. Chylomicrons were isolated lipids extracted and levels measured using commercial kits. N = 5 mice/group. PL, glycerophospholipids; C, total cholesterol. (D) Chylomicron size. Chylomicrons were isolated from wild-type and <i>Ces1/Es-x^−/−</i> mice and size was evaluated by dynamic light scattering at 25°C. N = 5 mice/group, 5 measurements per sample. (E) Chylomicron secretion. Overnight fasted mice were injected with P-407, followed by an olive oil bolus containing radiolabeled triolein. Blood was collected at the indicated times and plasma prepared. Lipids were extracted, spotted onto TLC plates and resolved. Lipids were visualized by exposure to iodine, and radioactivity in TG was counted in a scintillation counter. N = 6 mice/group, *</p

Primers used for quantitative real time PCR.

<p>Primers used for quantitative real time PCR.</p

Decreased expression of lipid absorption and increased expression of lipid secretion markers in <i>Ces1/Es-x^−/−</i> mice.

<p>(A) Intestinal expression of lipid absorption, synthesis and secretion genes in intestines from 24-week old mice was analyzed by qPCR. Mice were fasted for 4 h. N = 5 mice/group. *p<0.05 vs. wild-types. (B) Immunoblot of intestinal ApoB48. (D) Immunoblot of intestinal MTP. Same animals as in (A) were used. Cnx, calnexin  =  loading control. (D) and (E) quantitation of the immunoreactive ApoB48 and MTP bands, respectively. **p<0.01 vs. wild-type.</p

Lower TG accumulation in enterocytes from <i>Ces1/Es-x^−/−</i> mice.

<p>(A) and (B) Enterocytes from fasted mice were isolated and incubated with micelles containing radiolabeled oleic acid. Cellular radiolabel incorporation into TG and media radiolabeled TG secretion were assessed by lipid extraction, TLC separation, iodine exposure and scintillation counting. Cellular and media TG mass was measured by gas chromatography. N = 4 mice/group, *p<0.05, ***p<0.001. (C) Lipid content in intestinal mucosal scrapings from mice fasted overnight and re-fed for 6 h. Lipid standards in the last two lanes on the right are: DG, diacylglycerol (dioleoylglycerol); OA, oleic acid, TG, triacylglycerol (triolein), CE, cholesteryl ester (cholesteryl oleate). (D) Quantitation of the TG band in (C). (E) Lipid content in intestinal mucosal scrapings from mice fasted overnight. Lipid standards in the last two lanes on the right are as in (C). (F) Quantitation of the TG band in (E). *p<0.05 vs. wild-type.</p

Additional file 1: of Chemotherapy diminishes lipid storage capacity of adipose tissue in a preclinical model of colon cancer

Lipid metabolism proteins differentially expressed in tumour-bearing animals compared to the reference animals. (XLS 43 kb

Additional file 2: of Chemotherapy diminishes lipid storage capacity of adipose tissue in a preclinical model of colon cancer

Differentially expressed proteins in periuterine adipose tissue of animals receiving 2- cycles of chemotherapy compared to tumour-bearing animals. A total of 121 proteins were differentially expressed in periuterine adipose tissue after 2 cycles of chemotherapy compared to the tumour. (DOCX 58 kb