DMM Journal
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Disease Models & Mechanisms - discovery for human health. A
@biologists.bsky.social Open Access journal that publishes human disease research.
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![Microtubule mass is reduced in SDS patient cells. (A) mRNA levels of MAP7D1 in control and patient fibroblasts. The experiment was performed in three biological replicates, each including three independent technical replicates. qRT-PCR data in A represent the MAP7D1/ACTB values obtained by normalizing MAP7D1 mRNA expression levels in each experimental group to its respective ACTB mRNA expression level using the ΔΔCT method and compared with those of the control experiment group. (B) Quantification of endogenous MAP7D1 protein levels in control and patient fibroblasts. The expression level of endogenous MAP7D1 protein was analyzed by western blotting (left) with β-actin as an internal loading control. The experiment was performed in three biological replicates. Quantification of band densities is represented in the bar graph (right). Data were normalized to those of β-actin and compared with those of the control experiment group. (C) Confocal images showing colocalization of MAP7D1 protein with microtubules in control and patient fibroblasts (left and right panel, respectively) using immunocytochemistry. Antibody labeling for MAP7D1 is shown red, microtubules (stained for β-tubulin) are shown in green. Shown are representative of three biological replicates including two independent technical replicates. Merged images are shown on the right of each panel. Scale bars: 20 µm. (D) Quantification of the cell area in control and patient fibroblasts is represented in the bar graph (n≥45). (E) Quantification of the fluorescence intensity [measured as active fluorescent unit (AFU)] of microtubules in control and patient fibroblasts is represented in the bar graph (n≥50). t-test was used for all statistical analysis. Error bars represent the mean±s.e.m. *P<0.05, ***P<0.001, ****P<0.0001.](https://cdn.bsky.app/img/feed_thumbnail/plain/did:plc:5jupfg23qpjk2643o6aeftly/bafkreidk3z2fsmgpdnf6upqx23k2pbvvpf7xwtllbjvua7d43jg2xb3r3q@jpeg)
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![siCdk1 and siAnln decreased tumor burden in the DEN/PB HCC model. (A) Schematic overview of DEN/PB mouse model. At 2 weeks of age, male mice on the C3H strain background were given a single injection of DEN (25 mg/kg) 4 weeks before the start timepoint (-4w). At 6 weeks of age, mice were placed on 0.05% PB drinking water [start timepoint of 0 weeks (0w)]. Mice were injected with siRNA every other week starting at 12 weeks of age (6w), for a total of nine doses (with 5 mg/kg per dose) and sacrificed 1 week after the final dose (23w). (B) Liver weights for DEN/PB mice at 23 weeks after starting PB water, i.e. the final time point. (C) Liver-to-body-weight ratio for DEN/PB mice at the final time point. (D) Representative images of livers from DEN/PB mice at the final time point. (E) Surface tumor numbers were quantified using front and back images of each liver. (F) Representative immunofluorescence images of DEN/PB livers obtained from mice as indicated. Staining was against GS (red) to indicate the degree of WNT activation. Tumors are outlined by yellow dashed lines. One GS+ tumor was seen in the PBS group. Nuclei were stained with DAPI (blue). Scale bars: 800 μm. For panels B, C and E, the P-values with respect to the PBS group are shown above each group.](https://cdn.bsky.app/img/feed_thumbnail/plain/did:plc:5jupfg23qpjk2643o6aeftly/bafkreia346vromeksdvvhk4ikwhmd5jzwh35ijdqviiprwfwklidcshhqm@jpeg)
Reposted by DMM Journal
Reposted by DMM Journal
Reposted by DMM Journal