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Additional file 4: Fig. S4. Loss of either HCR or astrin slightly affects the protein level of CEP72. Parental HeLa cells, HCR-KO HeLa cells, and astrin-KO HeLa cells were analyzed by immunoblotting with CEP72 and beta-actin antibodies. Fig. S5. HCR does not bind to CEP63 and CEP152. HeLa cell lysates were immunoprecipitated with antibodies specific for HCR and negative control IgG. The precipitates were analyzed by immunoblotting with antibodies against CEP63 and CEP152. Fig. S6. Knockdown of astrin and HCR also caused microtubules organization defects in RPE cells. Microtubule regrowth assay of negative control, astrin and HCR siRNA-treated RPE cells co-stained with gamma-tubulin (green), alpha-tubulin (red), and DAPI (blue); scale bars, 10 μm. Fig. S7. Knockdown of astrin and HCR also caused mitotic spindle defects in RPE cells. Negative control, astrin and HCR siRNA-treated HeLa cells were treated with 100 ng/ml nocodazole for 16 hours to be arrested in M-phase and co-stained with gamma-tubulin (green), alpha-tubulin (red), and DAPI (blue); scale bars, 10 μm.

Additional file 9. Verification reports of astrin-KO cell line.

Additional file 5: Fig. S8. Co-localization of HCR with centrin1 and PCM1. HeLa cells transfected with HCR-GFP were co-stain with centrin1 (red) and gamma-tubulin (cyan) and DAPI (blue) or PCM1 (red) and gamma-tubulin (cyan) and DAPI (blue); scale bars, 10 μm. Fig. S9. HCR interacts with CEP131. HeLa cell lysates were immunoprecipitated with antibodies specific for HCR, astrin, or negative control rabbit IgG. The precipitates were analyzed by immunoblotting with antibodies against HCR and CEP131. Fig. S10. HCR did not interact with SKAP, MYCBP and LC8. HeLa cell lysates were immunoprecipitated with astrin, HCR, or control rabbit IgG antibodies and analyzed by western blotting with astrin, HCR, SKAP, MYCBP and LC8 antibodies. Beta actin antibody was used as negative control; HeLa cell lysates were immunoprecipitated with SKAP, MYCBP and LC8, or control rabbit IgG antibodies and analyzed by western blotting with astrin, HCR, SKAP, MYCBP and LC8 antibodies. Beta actin antibody was used as negative control. Fig. S11. HCR interacts with NuMA. mCherry vector alone or HCR-mCherry was transfected into HeLa cells for immunoprecipitation with mCherry antibody. The precipitates were analyzed by immunoblotting with HCR and NuMA antibodies. Fig. S12. HCR co-localizes with astrin and EDC4.HCR-GFP-transfected HeLa cells were treated with arsenite for 30 min and then co-stained with astrin (red) and EDC4 (cyan) for immunofluorescence detection; scale bars, 10 μm. Fig. S13. HCR co-localizes with EDC4 in mitosis. Negative control, HCR siRNA-treated HeLa cells were co-stained with HCR (red) and EDC4 (green); scale bars, 10 μm. Fig. S14. HCR was closely related to tumorigenesis. The dataset was from TCGA ( portal.gdc.cancer.gov ) and analyzed by TIMER 2.0 ( cistrome.org ).

Additional file 3: Fig. S3. Apoptosis or cycle changes in astrin-KO cells and identification of astrin-KO cell line. (A) Parental, astrin-KO and HCR-KO HeLa cells were analyzed with astrin, HCR, cyclin B1 and Cleaved PARP antibodies. HeLa cells treated with DMSO, Paclitaxel, Bafilomycin and MG132 were analyzed with Cleaved PARP and HCR antibodies. (B) Parental and astrin-KO HeLa cells were immunoblotted with an astrin antibody.

Additional file 1: Fig. S1. Repeated verification of the interaction between HCR and astrin. (A) Astrin Co-IP HCR and CEP72 and HCR Co-IP astrin and CEP72. HeLa cell lysates were immunoprecipitated with astrin, HCR, or control rabbit IgG antibodies and analyzed by western blotting with anti-astrin, anti-HCR antibodies. Anti-GM130 and anti-beta actin antibodies were used as negative control. The blotting of astrin, HCR and GM130 antibodies were incubated on the same membrane by repeatedly washing the membrane with antibody removal solution to increase comparability. (B) HCR interacts with astrin in HEK293 and U2OS cells. mCherry vector alone or HCR-mCherry in conjunction with the GFP-astrin plasmid were transfected into HEK293 cells for immunoprecipitation using an mCherry antibody. The eluted proteins were analyzed with mCherry and GFP antibodies. HCR-mCherry plasmid-transfected HEK293 cells were immunoprecipitated with astrin antibody or negative control rabbit IgG. The precipitates were analyzed with mCherry and astrin antibodies. U2OS cell lysates were immunoprecipitated with an HCR antibody or negative control rabbit IgG. The precipitates were analyzed with antibodies against HCR and astrin.

Additional file 2: Fig. S2. Identification of HCR-GFP stable Cell Line and Localization of HCR in cells. (A) Identification of HCR-KO HeLa cell line and stably expressing HCR-GFP cell line. Parental HeLa cells, HCR-KO HeLa cells, HCR-KO cells transfected with HCR-GFP, and stably transfected HCR-GFP HCR-KO cells were immunoblotted with an HCR antibody. (B) Co-localization of astrin-CC2 and HCR-CC3. HeLa cells transfected with astrin-CC2-myc and HCR-GFP or astrin-CC2-myc and HCR-CC3-GFP were co-stained with myc (red) and gamma-tubulin (cyan); scale bars, 10 μm. (C) Co-localization of HCR with alpha-tubulin. Mitotic HeLa cells stained with an alpha-tubulin antibody (green), HCR antibody (red), and DAPI (blue) for nuclear staining (left panel) or stained with anti-alpha-tubulin (green), anti-astrin (red), and DAPI (blue) (right panel); scale bars, 10 μm. (D) Identification of antibody staining to HCR. Negative control, HCR siRNA-treated HeLa cells were co-stained with HCR (red) and alpha-tubulin (green); scale bars, 10 μm. (E) The effect of Nocodazole on HCR is dose-dependent and recoverable. HeLa cells were treated with 1μM, 0.75μM, 0.5μM Nocodazole for 5 hours or treated with 1μM Nocodazole for 5hours then released from Nocodazole for 30 min, 1 hour, 2 hours, then co-stained with HCR (red) and gamma-tubulin (green). (F) Knockdown of HCR does not affect PCM1 localization. Negative control, HCR siRNA-treated HeLa cells were co-stained with HCR (red) and PCM1 (green); scale bars, 10 μm.

After pipeline construction burst china energy pipeline industry commonly faced with aging and abandonment problem. The study on pipeline abandonment was still in preliminary state, because there is nearly no pipeline abandonment standards and guidelines. Pipeline cleaning is the first step to dispose the abandoned pipeline to eliminate the risk of environment and safety. In foreign developed countries there are many chemical cleaning agents. But this chemical cleaning agent is not suitable for residual of chines pipeline because of oil difference between china and foreign country. The most of residual in china have a very high paraffin and resin-asphaltenes content. So preparation of chemical cleaning suitable for china residual is a challenge on the technology of abandonment pipeline. In this paper two kinds of chemical cleaning agent were prepared to clean the different type of residual. They are hydrophilic chemical cleaning agent and oil soluble chemical cleaning agent. The result of cleaning project shows that the both kind of chemical cleaning have good performance for the abandoned pipeline. The chemical cleaning agents will assist pipeline company to totally clean the abandoned pipeline to ensure the public safety and environment protection.

China crude oil pipeline industry commonly faces with abandonment problem because of the decreasing output of oilfield and pipeline integrity problem. China released its first standard on abandoned pipeline (SY/T 7413-2018) named specifications for the disposal of abandoned oil&gas pipelines and it is implemented on the April 1st 2019. Pipeline cleaning is very necessary to dispose the abandoned pipeline to eliminate the risk of environment and safety. There are many cleaning agents for abandoned pipeline in the market. This chemical cleaning agent is not suitable for china pipeline because the wax remained in china pipeline is different that of foreign pipeline. The wax content in residual of china pipeline is very high. So, preparation of cleaning agent suitable for china pipeline is very important to remove wax with low cost. In this study one kind of hydrophilic cleaning agent is prepared. The result of lab test result and cleaning project both show that the kind of cleaning have excellent performance for the abandoned pipeline.

On the basis of the study on the principle of oil flow and that on mechanism of drag reduction and transportation promotion of oil pipelines, the article makes a further research on the Mechanism of Drag Reduction in the gas pipelines. It points out that the basic cause for gas pipelines drag reduction is to control effectively the radial pulsation of gas adjacent to pipeline wall. It is considered that the most effective drag reduction method is to reduce the pipeline wall’s roughness degree and pipeline wall’s undercoat and to make gas drag reduction agent adjacent to pipeline wall. The research shows that gas drag reduction agent should be a polymer and compound with polarity and nonpolarity long chain.Copyright © 2008 by ASME