General information of subjects before drug switch.

<p>General information of subjects before drug switch.</p

Resistance to 3TC and TDF before and after drug switch.

<p>Resistance to 3TC and TDF before and after drug switch.</p

CD4 count, viral load and viral inhibition rates before, 6 months after, and 12 months after drug switch.

<p>CD4 count, viral load and viral inhibition rates before, 6 months after, and 12 months after drug switch.</p

Drug resistance analysis before and after second-line treatment.

<p><b>*</b>A score >60 get from Stanford University drug resistance database was regarded as high drug resistance.</p><p>Drug resistance analysis before and after second-line treatment.</p

HPV16 E6 binds to ING4 and hinders its association with p53.

<p>(A) HPV16 E6 does not affect ING4 stability. p53^−/−Mdm2^−/−MEF cells were transfected with ING4 and an increasing amount of either Flag-HPV16 E6 or control vector. The levels of ING4, HPV16 E6 and GAPDH were examined by Western blot. (B) HPV16 E6, ING4 and p53 bind each other. Flag-E6, PCDNA-ING4, and Myc-p53 were transfected into p53^−/−Mdm2^−/−MEF cells. Cell lysates were immunoprecipitated with EzviewTM Red anti-Flag M2 Affinity Gel (lanes 3 and 4), Flag-E6 and the associated proteins were eluted with 3XFlag peptide. Twenty percent of the eluent was subject to Western analysis using indicated antibodies. The remaining eluent was used for secondary immunoprecipitation with anti-myc antibody (lanes 5and 6). (C) Saos2 cells were transfected with PCDNA-ING4, Myc-p53, and either increasing amounts of HPV16 E6 or the vector control. Transfected cells were treated with MG132 for 6 h. The association of PCDNA-ING4 and Myc-p53 was analyzed by immunoprecipitation assay with anti-p53 antibody. (D) Knockdown of HPV16 E6 increases the ING4-p53 interaction. CaSki and SiHa cells with lentivirus-delivered GFP labeled shRNA against HPV16 E6 (shE6) or scramble control (shC) were shown on the left panel. On the other side, the top panel showed the whole cell lysates were immunoprecipitated with anti-p53 antibody and checked ING4 expression by Western blot. Input equivalent to 10% of the whole cell lysates used for immunoprecipitation was subjected to Western blot using the indicated antibody. (E) ING4 mediated p53 acetylation was attenuated by HPV16 E6. Saos2 cells were transfected with PCDNA-ING4, Myc-p53, in the present of either HPV16 E6 or its mutant L50G. At 24 hr post transfection, trichostatin A were added for additional 6 hours, immunoprecipitation and western blot showed that HPV16 E6L50G attenuated binding and acetylation induced by ING4 on p53 without p53 degradation mediated by E6AP. (F) HPV16 E6 attenuates the binding affinity between ING4 and p53 in vitro. Myc-tagged p53 was incubated with bacterially-expressed GST or GST-ING4 in the presence of Flag-tagged HPV16 E6 or control vector for GST-pull down assay. Concentration of each fusion protein used in GST-pull down assays was kept the same. The level of myc-p53 pulled down by GST-ING4 was less in the presence of HPV16 E6.</p

HPV16 E6 forms complex with ING4 independent of p53.

<p>(A) Lysates from HPV negative cervical carcinoma cell line C33A and two HPV16 positive cell lines CaSki and SiHa were subjected to IP with HPV16 E6 specific antibody C1P5 and detected by Western blotting (WB) for the indicated proteins. (B) Saos2 cells were co-transfected with Flag-tagged E6 and PCDNA-ING4, balanced with empty vector. The cell lydates were subjected to IP with ING4 specific antibody and detected by WB. (C) and (D) Either GST control, GST-ING4 full length or truncates beads were incubated with HPV16 E6 in vitro translated protein with ³⁵S-radiolabeled. 5% of in vitro translated protein input was used as a comparison. Precipitated proteins were resolved by SDS-PAGE, exposed to phosphorimager screen and scanned by Typhoon 9410 imaging system. Coomassie blue staining of SDS-PAGE-resolved purified GST and GST-ING4 proteins was shown under the panel. (E) Colocalization of ectopically expressed ING4 and HPV16 E6. Saos2 cells plated on coverslips were transfected with Flag-E6 and PCDNA-ING4 using Lipofectamine 2000. (F) Colocalization of endogenous HPV16 E6 and ING4 in CaSki cells. All panels are representative pictures from approximately 50 cells of five different fields of three independent experiments.</p

HPV16 E6 suppresses the ING4 mediated p53 transcriptional activity and apoptosis.

<p>(A ) Saos-2 (p53^−/−) cells were cotransfected with a wild-type p21WAF1/CIP1 promoter construct, different combinations of plasmids expressing Myc-p53, PCDNA-ING4, flag-tagged HPV16 E6 or its mutant L50G. At 36 h posttransfection, cells were harvested and lysed in reporter lysis buffer. The bars plot the means of the results of two independent experiments. Error bars represent standard deviations (SD). The results showed that HPV16 E6 attenuates ING4-triggered p53 transcriptional activity. The expression levels of each target proteins were detected by western blotting and shown at the bottom panels. (B) Saos2 cells were transfected with expression plasmids for myc-p53, PCDNA-ING4, in combinations with flag-E6 or its mutant L50G. After a 2-week selection, cells were fixed on the plates with 4% formaldehyde and stained with 0.1% crystal violet. A representative of colony formation was shown. The area of colonies (pixels) in each dish was calculated by Li-Cor Odyssey. The number represents the averages of data from two independent experiments.</p

HPV16 E6 suppresses the ING4 mediated p53 transcriptional activity and apoptosis.

<p>(C) U2OS cells were respectively transfected with siRNA Luciferase or siRNA E6AP, with ING4 or ING4+E6 using the Lipofectamin 2000. Cells were collected at 36 h posttransfection after a 12-h serum starvation and fixed. Levels of cells undergoing apoptosis (sub-G1 phase) in individual PI-stained samples were analyzed by flow cytometry, and the data were analyzed by FlowJo software. The bar diagram represents the mean of three independent experiments. Western blot showing the protein level of E6AP in the lentivirus-mediated E6AP of control knockdown cell lines. GAPDH was used as the loading control.</p

Ultrafastly Interweaving Graphdiyne Nanochain on Arbitrary Substrates and Its Performance as a Supercapacitor Electrode

A moderate method is first developed here for superfast (in seconds) growth of an ultrafine graphdiyne (GDY) nanochain on arbitrary substrates in the atmosphere. This is an environmentally friendly and metal-catalyst-free method, efficiently eliminating extraneous contaminations for the carbon materials. The seamless GDY coating on any substrates demonstrates that an all-carbon GDY possesses outstanding controllability and processability, perfectly compensating for the drawbacks of prevailing all-carbon materials. After the decoration of 3D GDY nanostructures, the substrates become superhydrophobic with contact angles high up to of 148° and can be used as outstanding frameworks for storing organic pollution. Because of the reasonable porous and 3D continuous features, the as-prepared samples can be applied as high-performance binder-free supercapacitor electrodes with high area capacitance of up to 53.66 mF cm ^–2, prominent power performance, and robust long-term retention (99% after 1300 cycles)

Molecular characteristics of all hVISA and VISA isolates.

*<p>NT, could not be typed.</p