Journal: Journal of Biological Chemistry

Article Title: Atypical Protein Kinase Cι Plays a Critical Role in Human Lung Cancer Cell Growth and Tumorigenicity

doi: 10.1074/jbc.m505402200

Figure Lengend Snippet: FIG. 2. PKC does not regulate NFB signaling in A549 cells. A, immunoblot analysis of A549 cell transfectants for cIAP2, Bcl-XL, and actin. B, analysis of A549 cell transfectants for PARP and cleaved PARP. HeLa cells treated with taxol for either 24 or 48 h served as a positive control for cleaved PARP. C, A549/pBabe and A549/kdPKC cells were treated with 50 g/ml TNF for the indicated times and cell lysates subjected to immunoblot analysis for detection of IB, phospho-(Ser32)-IB, and actin. D, A549/pBabe and A549/kdPKC cells were treated with 50 g/ml TNF for the indicated times and cell lysates subjected to immunoblot analysis for detection of NFB p65, phospho-(Ser536)-NFB, p65, and actin. E, transcriptional activity of an NFB-luciferase reporter in A549/pBabe and A549/kdPKC cells in the presence and absence of TNF. Data represent the mean S.E. of three independent determinations.

Article Snippet: This paper is available on line at http://www.jbc.org 31109 at K U N G L IG A T E K N ISK A H O G SK O L A N on M arch 8, 2015 http://w w w .jbc.org/ D ow nloaded from (Sigma, catalog number A8592); Rac1, 1:3000 (BD Transduction Laboratories, catalog number 610651); cIAP2, 1:500 (Santa Cruz Biotechnology, catalog number sc-7944); Bcl-XL, 1:1,000 (Cell Signaling Technologies, catalog number 2762); PARP/cleaved PARP, 1:1,000 (Cell Signaling Technologies, catalog number 9542); I B , 1:1,000 (Cell Signaling Technologies, catalog number 9242); phospho-(Ser32)-I B , 1:1,000 (Cell Signaling Technologies, catalog number 9241); NF B p65, 1:1,000 (Cell Signaling Technologies, catalog number 3034); phospho(Ser536)-NF B p65, 1:1,000 (Cell Signaling Technologies, catalog number 3031); Mek, phospho-(Ser217/221)-Mek, and phospho-(Ser298)Mek, each at 1:1,000 (Cell Signaling Technologies, catalog numbers 9122, 9121, and 9128, respectively), Erk and phospho-(Thr202/Tyr204)Erk, each at 1:1,000 (Cell Signaling Technologies, catalog numbers 9102 and 9101, respectively); CD31, 1:1,000 (or Pecam-1; Santa Cruz Biotechnology, catalog number sc-1506); BrdUrd, 1:100 (DAKO, catalog number M0744).

Techniques: Western Blot, Positive Control, Activity Assay, Luciferase

Journal: iScience

Article Title: Cancer cell intrinsic TIM-3 induces glioblastoma progression

doi: 10.1016/j.isci.2022.105329

Figure Lengend Snippet:

Article Snippet: NF-κB p65(S536) (93H1) , Cell Signaling Technology , 3033S; RRID: AB_331284.

Techniques: Control, Purification, Recombinant, Modification, Sterility, Cell Culture, Lysis, Transfection, Western Blot, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Proliferation Assay, Imaging, Sequencing, Real-time Polymerase Chain Reaction, Plasmid Preparation, Software, Fluorescence, Microscopy

Journal: Experimental and therapeutic medicine

Article Title: HIPK2 sustains inflammatory cytokine production by promoting endoplasmic reticulum stress in macrophages.

doi: 10.3892/etm.2020.9301

Figure Lengend Snippet: Figure 2. HIPK2 knockdown suppresses IL‑6 and TNF‑α production in macrophages. (A) Knockdown efficacy of si‑HIPK2 and controls at the mRNA level in LPS‑stimulated PMs at the indicated time points (n=3). (B) Western blot analysis of HIPK2 following siRNA interference, and levels of p‑p65 and p65 in PMs (three independent repeated experiments were performed). ELISA analysis of (C) IL‑6 and (D) TNF‑α in the supernatant following HIPK2 knockdown in LPS‑stimulated PMs (n=4). *P<0.05 vs. control; #P<0.05 vs. corresponding si‑controls in the same group. HIPK2, homeodomain‑interacting protein kinase 2; si, small interfering RNA; PMs, peritoneal macrophages; p‑, phosphorylated; LPS, lipopolysaccharides.

Article Snippet: The primary antibodies used in the present study were: Anti‐mouse HIPK2 (cat. no. 5091; Cell Signaling Technology, Inc.), phosphorylated (p‐) p65 (cat. no. 3033; Cell Signaling Technology, Inc.), p65 cat. no. 9460; Cell Signaling Technology, Inc.), phosphorylated (p‐)PERK (cat. no. 3179S; Cell Signaling Technology, Inc.), PERK (cat. no. 3192S; Cell Signaling Technology, Inc.), p‐IRE1α (cat. no. ab48187; Abcam), IRE1α (cat no. 3294S; Cell Signaling Technology, Inc.), ATF6 (cat. no. ab122897; Abcam) and β‐actin (cat. no. ab8227; Abcam).

Techniques: Knockdown, Western Blot, Enzyme-linked Immunosorbent Assay, Control, Small Interfering RNA

Journal: Experimental and therapeutic medicine

Article Title: HIPK2 sustains inflammatory cytokine production by promoting endoplasmic reticulum stress in macrophages.

doi: 10.3892/etm.2020.9301

Figure Lengend Snippet: Figure 4. Activation of endoplasmic reticulum stress restores IL‑6 and TNF‑α production following HIPK2 knockdown in LPS‑stimulated macrophages. ELISA analysis of (A) IL‑6 and (B) TNF‑α levels in the supernatants with or without TM treatment in PMs following HIPK2 knockdown (n=3). (C) Western blot analysis of p‑p65 and p65 levels in PMs treated with or without TM following HIPK2 knockdown. *P<0.05 as indicated. HIPK2, homeodomain‑interacting protein kinase 2; LPS, lipopolysaccharides; PMs, peritoneal macrophages; TM, tunicamycin; p‑, phosphorylated; si, small interfering RNA; NS, not significant.

Article Snippet: The primary antibodies used in the present study were: Anti‐mouse HIPK2 (cat. no. 5091; Cell Signaling Technology, Inc.), phosphorylated (p‐) p65 (cat. no. 3033; Cell Signaling Technology, Inc.), p65 cat. no. 9460; Cell Signaling Technology, Inc.), phosphorylated (p‐)PERK (cat. no. 3179S; Cell Signaling Technology, Inc.), PERK (cat. no. 3192S; Cell Signaling Technology, Inc.), p‐IRE1α (cat. no. ab48187; Abcam), IRE1α (cat no. 3294S; Cell Signaling Technology, Inc.), ATF6 (cat. no. ab122897; Abcam) and β‐actin (cat. no. ab8227; Abcam).

Techniques: Activation Assay, Knockdown, Enzyme-linked Immunosorbent Assay, Western Blot, Small Interfering RNA