Journal: Acta Pharmaceutica Sinica. B

Article Title: A new perspective of triptolide-associated hepatotoxicity: the relevance of NF- κ B and NF- κ B-mediated cellular FLICE-inhibitory protein

doi: 10.1016/j.apsb.2020.02.009

Figure Lengend Snippet: Possible role of NF- κ B and NF- κ B-mediated pro-survival signals in TP/LPS-induced hepatotoxicity. (A) Experimental design to detect the time-dependent changes of NF- κ B and its related pro-survival factors. (B)–(G) Relative mRNA levels of NF- κ B target genes, including Ciap1 , Ciap2 , Flip L , Xiap , Tnfaip3 , and Nfkbia , were detected by qPCR with tubulin as the internal control ( n = 6). (H)–(L) Representative Western blots and relative intensity of protein bands of FLIP L , CIAP1, XIAP, and I κ B- α after the treatment of TP and LPS with tubulin as the loading control ( n = 4–6). (M) Representative photomicrographs of liver sections by IHC for P65 1 h after LPS application (200 × ). Scale bar = 50 μm. Results were expressed as mean ± SEM and statistical analysis was performed using Two-way ANOVA following by Tukey's multiple comparison test. *, # , $ P <0.05, **, ## , $$ P <0.01, ***, ### , $$$ P <0.001; ns, no statistical difference.

Article Snippet: Antibody against RIPK1 (17519-1-AP), mixed lineage kinase domain like pseudokinase (MLKL, 66675-1-Ig), myeloperoxidase (MPO, 22225-1-AP), and CIAP1 (10022-1-AP) were purchased from Proteintech (Chicago, IL, USA).

Techniques: Control, Western Blot, Comparison

Journal: Acta Pharmaceutica Sinica. B

Article Title: A new perspective of triptolide-associated hepatotoxicity: the relevance of NF- κ B and NF- κ B-mediated cellular FLICE-inhibitory protein

doi: 10.1016/j.apsb.2020.02.009

Figure Lengend Snippet: The primer sequences used for qPCR assay in mice.

Article Snippet: Antibody against RIPK1 (17519-1-AP), mixed lineage kinase domain like pseudokinase (MLKL, 66675-1-Ig), myeloperoxidase (MPO, 22225-1-AP), and CIAP1 (10022-1-AP) were purchased from Proteintech (Chicago, IL, USA).

Techniques:

Journal: Journal of Biological Chemistry

Article Title: Cellular Inhibitor of Apoptosis Protein 1 (cIAP1) Stability Contributes to YM155 Resistance in Human Gastric Cancer Cells

doi: 10.1074/jbc.m114.600874

Figure Lengend Snippet: FIGURE 3. Double knockdown of survivin and cIAP1 induces cell death in YM155-resistant MKN45 cells. A and B, KATOIII (A) and MKN45 (B) cells were co-transfected with survivin-siRNA and cIAP1 siRNA for 48 h, and then cell death was determined using the trypan blue exclusion method. Cell lysates were subjected to Western blot analysis with anti-survivin and anti-CIAP1 and the loading control -actin. The data are the mean S.D.; **, p 0.01.

Article Snippet: Plasmids, siRNAs, and Transfections—Survivin and cIAP1 cDNAs were purchased from Origene (Rockville, MD).

Techniques: Knockdown, Transfection, Western Blot, Control

Journal: Journal of Biological Chemistry

Article Title: Cellular Inhibitor of Apoptosis Protein 1 (cIAP1) Stability Contributes to YM155 Resistance in Human Gastric Cancer Cells

doi: 10.1074/jbc.m114.600874

Figure Lengend Snippet: FIGURE 6. The survivin heteromer decreases after YM155 in YM155-sensitive KATOIII cells but not MKN45 cells. A, MKN45 cells were immunoprecipi- tated with anti-survivin or anti-cIAP1 or anti-rabbit IgG antibodies. Interaction of survivin and cIAP1 was determined with Western blot analysis using anti-survivin or anti-cIAP1 antibodies. B, KATOIII and MKN45 cells were treated with YM155 (20 nM) for 48 h. Cell lysates were run on a native gel. Various heteromers of survivin were determined with Western blot analysis using anti-survivin antibody. C, KATOIII and MKN45 cells were treated with YM155 (20 nM) for 48 h and then cell lysates were immunoprecipitated (IP) using an anti-survivin antibody. Determination of the binding of survivin to the monomer or dimer of cIAP1 was determined via native PAGE using anti-cIAP1.

Article Snippet: Plasmids, siRNAs, and Transfections—Survivin and cIAP1 cDNAs were purchased from Origene (Rockville, MD).

Techniques: Western Blot, Immunoprecipitation, Binding Assay, Clear Native PAGE