Deadend colorimetric tunel detection kit

A total volume of 200 μl of 5-8F/shc-Myc and 5-8F/shCtrl cells (1× 10⁷ cells) transfected with miR-141 mimic (miR-141) or negative control (miR-NC) was inoculated subcutaneously into the left flanks of 6-week-old female nude mice. Mice were checked every 4 days. Tumor volume was evaluated using the following formula: volume = (width + length)/2 × width × length × 0.5236. All three groups were killed after 28 days. All tumor grafts were excised, weighed, harvested, fixed and embedded. The anti-Ki67 antibody (dilution 1:100, Bioworld) was used to detect the proliferation marker Ki67 using IHC procedures, and a TUNEL assay was used to detect apoptosis in situ with paraffin-embedded xenograft sections using the DeadEnd Colorimetric TUNEL Detection Kit (Promega) according to the manufacturer’s protocols. Samples were observed using an Olympus microscope (Olympus, Tokyo, Japan). The proliferative and apoptosis index scores were measured as the mean percentage of nuclei that stained positive for Ki67 and TUNEL cells in 10 different 200× fields.

A total volume of 100 μl of cells (5 × 10⁶ cells) transfected with miR-141 mimic or negative control were inoculated subcutaneously into the right flanks of 6-week-old female nude mice. Mice were checked every 4 days, and tumor nodules were measured with a caliper. Tumor volume was evaluated using the following formula: volume=(width+length)/2 × width × length × 0.5236. Tumor growth curves were calculated. The three experimental groups were killed after 32 days. All tumor grafts were excised, weighed, harvested, fixed and embedded. The anti-Ki-67 antibody (dilution 1:100, Bioworld) was used to detect the proliferation marker Ki-67 using immunohistochemistry procedures, and TUNEL assay was used to detect apoptosis in situ with paraffin-embedded xenograft sections using the DeadEnd Colorimetric TUNEL Detection Kit (Promega) according to the manufacturer's protocols. Samples were observed using an Olympus microscope (Olympus, Tokyo, Japan). The proliferative and apoptosis index scores were measured as the mean percentage of nuclei that stained positive for Ki-67 and TUNEL cells in 10 different × 200 fields.