Journal: Cell Reports

Article Title: Modeling of Cisplatin-Induced Signaling Dynamics in Triple-Negative Breast Cancer Cells Reveals Mediators of Sensitivity

doi: 10.1016/j.celrep.2019.07.070

Figure Lengend Snippet:

Article Snippet: Most antibodies used in this study were purchased from Cell Signaling, including those targeting γH2AX (#9718), CDC25C (#4688), p-CHK1-Ser345 (#2348), p-CHK2-Thr68 (#2197), p-MK2-Thr334 (#3041), p-ATR-Ser428 (#2853), p-AKT-Ser473 (#3787), p-ERK-Thr202/Tyr204 (#4376), p-P38-Thr180/Tyr182 (#4511), p-JNK-Thr183/Tyr185 (#9251), p-NFκB p65-Ser536 (#3033), RIPK1 (#3493), Aurora A (#3092), Bcl-xL (#2762) and MCL1 (#4572).

Techniques: Recombinant, Protease Inhibitor, Blocking Assay, Expressing, Bradford Protein Assay, Inhibition, Software

Journal: Oncology reports

Article Title: NF-κB activity is downregulated by KRAS knockdown in SW620 cells via the RAS-ERK-IκBα pathway.

doi: 10.3892/or.2012.1669

Figure Lengend Snippet: Figure 1. (A and B) Constitutive activation of NF-κB in colon cancer cells. Cytoplasmic and nuclear lysates were analyzed by western blot analysis. β-actin (cytoplasmic) and PARP (nuclear) were used to determine purity. (C) Decreased NF-κB activation in KRAS knockdown SW620 cells. SW620 cells were transfected by KRAS shRNA or control shRNA lentiviral particles. RAS (total cellular lysates) and p65 (nuclear lysates) were analyzed by western blot analysis. β-actin (cytoplasmic) and PARP (nuclear) were used as a loading control. Line 1: negative control, Line 2: control shRNA, Line 3: KRAS shRNA. (D) U0126 downregulated nuclear p65 and phosphorylation of ERK, and IκB in SW620 cells. Cells were treated with 3 or 10 µM U0126 and collected after 30 min, followed by immunoblotting using anti-p65, anti-phospho-ERK or -IκB antibodies. β-actin (cytoplasmic) and PARP (nuclear) were used as a loading control. (E) Decreased NF-κB activation in KRAS knock-down SW620 cells probably through the RAS-ERK-IκBα pathway. SW620 cells were transfected by KRAS shRNA or control shRNA lentiviral particles. Cell extracts were immunoblotted to detect the indicated protein species. Line 1: negative control, Line 2: control shRNA, Line 3: KRAS shRNA.

Article Snippet: JHS-23 (20), a selective inhibitor of nuclear translocation of NF-κB p65, was purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Activation Assay, Western Blot, Knockdown, Transfection, shRNA, Control, Negative Control, Phospho-proteomics

Journal: Oncology reports

Article Title: NF-κB activity is downregulated by KRAS knockdown in SW620 cells via the RAS-ERK-IκBα pathway.

doi: 10.3892/or.2012.1669

Figure Lengend Snippet: Figure 4. Immunohistochemical positive staining for the nuclear factor-κB subunit p65 in primary colorectal cancer. Positivity of the tumor for NF-κB expression was defined as distinct nuclear immunostaining.

Article Snippet: JHS-23 (20), a selective inhibitor of nuclear translocation of NF-κB p65, was purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

Techniques: Immunohistochemical staining, Staining, Expressing, Immunostaining

Journal: Human Reproduction (Oxford, England)

Article Title: Periconceptional exposure to lopinavir, but not darunavir, impairs decidualization: a potential mechanism leading to poor birth outcomes in HIV-positive pregnancies

doi: 10.1093/humrep/deaa151

Figure Lengend Snippet: Lopinavir treatment inhibits the transcription factor STAT3. ( A ) Representative images of phosphorylated signal transducer and activator of transcription 3 (p-STAT3) immunostaining of stroma cells in first-trimester human decidua treated with DMSO (control) or protease inhibitor (PI)-combination antiretroviral therapy (cART) as indicated. Inset is negative control. Scale bar, 50 µm. ( B ) Quantification of staining intensity (pixels) of p-STAT3 positive stroma cells. Results are the mean ± SD. *** indicates P < 0.0001, calculated using ANOVA with Tukey’s post hoc analysis. Each data point represents the average of two to three replicates per experiment; n = 10 independent experiments. ( C ) Representative images of p-STAT3 immunostaining of GD 6.5 implantation sites of control, ritonavir-boosted lopinavir (LPV/r)-cART and ritonavir-boosted darunavir (DRV/r)-cART treated mice. E, embryo; PDZ, primary decidual zone; SDZ, secondary decidual zone; M, mesometrium; AM, anti-mesometrium. * indicates uterine lumen. Scale bar, 600 µm. ( D ) Quantification of staining intensity (pixels) of p-STAT3 positive cells. Results are the mean ± SD. *** indicates P < 0.0001, calculated using ANOVA with Tukey’s post hoc analysis. Each data point represents an implantation site, three implantation sites were analyzed per mouse (n = 4 mice/treatment group).

Article Snippet: Total STAT3 was measured in decidual cell lysates (n = 8) using the human STAT3 ELISA kit (Abcam ab176666) as per the manufacturer’s protocol.

Techniques: Immunostaining, Protease Inhibitor, Negative Control, Staining

Journal: bioRxiv

Article Title: RANK is an independent biomarker of poor prognosis in estrogen receptor-negative breast cancer and a therapeutic target in patient-derived xenografts

doi: 10.1101/2021.12.13.470911

Figure Lengend Snippet: (a) RANK and RANKL mRNA expression levels relative to PPIA in the indicated BC PDXs, organized according to ER status in the human tumor of origin and RANK mRNA expression. Two tailed t-student test was used to evaluate the RANK/RANKL differential expression between ER - and ER + BC PDXs. # Indicates models where RANK and RANKL expression were analyzed by IHC. (b) Percentage of PDXs expressing RANK protein according to ER expression. Total number of independent PDXs analyzed is shown. P-value was calculated using a two tailed t-student test. (c) Representative images of RANK and RANKL protein expression in BC PDXs detected by IHC. H-Score (H) of the models (and not of the picture) are shown. A total of 3-5 independent tumors per PDX, were scored for RANK. Pictures are ordered according to RANK mRNA expression levels and subtype in the human samples of origin. (d) Western blot analyses of P-p65, P-IKBα and corresponding total proteins after the minutes (min) of RANKL stimulation in the indicated PDXs. Tubulin was used as a loading control.

Article Snippet: Primary antibodies against P-p65 (Ser536, Cell Signaling), p65 (D14E12, Cell Signaling), P-IkBα (S32/36, Cell Signaling), IkBα (L35A5 Cell Signaling), and β-tubulin (ab21058, Abcam) were used.

Techniques: Expressing, Two Tailed Test, Quantitative Proteomics, Western Blot, Control