Journal: eLife

Article Title: The long noncoding RNA lnc-FANCI-2 intrinsically restricts RAS signaling in human papillomavirus type 16-infected cervical cancer cells

doi: 10.7554/eLife.102681

Figure Lengend Snippet: ( A ) The lnc-FANCI-2-associated proteins in the wild-type (WT) CaSki cells were identified by i solation of R NA- p rotein c omplexes using R NA p urification (IRPCRP)-mass spectrometry technology. ( B ) lnc-FANCI-2 RNA in the IRPCRP-1 and IRPCRP-2 pull-downs had the pooled antisense biotinylated oligos (pool 1 with oligos in even numbers and pool 2 with oligos in odd numbers) immobilized to avidin-beads first before mixed with cell lysates, while the IRPCRP-3 and IRPCRP-4 pull-downs had the oligos pool 1 and 2 separately mixed with cell lysates first before addition to the avidin-beads for the RNA pull-downs. RT-PCR in the absence (-) or presence (+) of reverse transcriptase (RT) was carried out using the RNA isolated from the individual IRPCRP experiments using a primer pair of oHBL5 and oHBL12 specific for lnc-FANCI-2 RNA detection. Beads-only (no oligos) IRPCRP experiments served as a negative control. Total RNA from the WT CaSki cells after sonication was used as an input control. The arrow indicates the detected lnc-FANCI-2 RNA. ( C ) Proteins associated with lnc-FANCI-2 RNA identified from lnc-FANCI-2 IRPCRP pull-downs. A total of 32 proteins were specifically pulled down from lnc-FANCI-2 IRPCRP reactions 1–4 (PSM ≥2 from two separate pull-downs, ), with top 10 proteins binding lnc-FANCI-2 shown in the order by the number of identified peptide spectrum match (PSM) in LC-MS/MS. ( D ) Expression of p-Akt and p-Erk from CaSki WT cells 48 hr after siRNA KD of MAP4K4 or lnc-FANCI-2 was immunoblotted by the corresponding antibodies. GAPDH served as a sample loading control. Fold change of the indicated proteins in the cells with KD of MAP4K4 or lnc-FANCI-2 over the cells treated by a non-targeting siRNA (siNS) was calculated after normalizing to GAPDH. ( E ) A proposed model illustrates how lnc-FANCI-2-protein complexes inhibit the RAS signaling pathway to control Akt/Erk phosphorylation and expression of host genes. RAS signaling can be regulated by integrating external and internal factors. In high-risk human papillomavirus (HR-HPV) infected cells, viral oncoprotein E7-YY1 complex transactivates lnc-FANCI-2 expression. By interactions with cellular RBPs, the lnc-FANCI-2-protein complex inhibits RAS signaling. In the absence of lnc-FANCI-2, increased RAS signaling leads to phosphorylation of Akt and Erk and cascaded responses of transcription factors (TFs) and thus regulates the expression of RAS signaling responder genes, such as IGFBP3, MCAM, etc. Consequently, this brings fundamental biochemical and biological processes under control by fine-tuning the RAS pathway. Figure 9—source data 1. lnc-FANCI-2 interacts with host factors to regulate RAS signaling pathway. Figure 9—source data 2. lnc-FANCI-2 interacts with host factors to regulate RAS signaling pathway.

Article Snippet: Total ribo-minus RNA-seq libraries, four samples in each group, were prepared with TruSeq Stranded Total RNA Library Kit and then subjected to paired-end sequencing using Illumina-HiSeq3000/4000 platform.

Techniques: Mass Spectrometry, Avidin-Biotin Assay, Reverse Transcription Polymerase Chain Reaction, Reverse Transcription, Isolation, RNA Detection, Negative Control, Sonication, Control, Binding Assay, Liquid Chromatography with Mass Spectroscopy, Expressing, Phospho-proteomics, Infection

Journal: Journal of Cellular and Molecular Medicine

Article Title: Fibroblast growth factor 2 protects against renal ischaemia/reperfusion injury by attenuating mitochondrial damage and proinflammatory signalling

doi: 10.1111/jcmm.13203

Figure Lengend Snippet: FGF2 inhibits I/RI‐induced HMGB1 serum release and inflammatory response. Animals were divided into 5 groups ( n = 4), including sham‐operated control, I/RI group, and I/RI with FGF2 pre‐treatment or delayed treatment at 1 and 12 hrs, respectively, after reperfusion as indicated. The samples were collected at 48 hrs following reperfusion for Western blot, ELISA, Immunohistochemistry staining (IHC) and qRT‐PCR analysis as detailed below. ( A ) Western blot analysis to determine the expression of HMGB1 and TNFα in renal tissues with GAPDH as loading control. ( B ) ELISA assay was used to determine the levels of HMGB1 in the serum of animals receiving indicated treatments. ** P < 0.01 versus sham group, ## P < 0.01 versus I/R group. ( C ) IHC of kidney tissue sections for expression of HMGB1. Original magnification ×20. One representative area of renal tissue staining from 1 of 4 animals in each group is shown. (D) Real‐time PCR quantification of mRNA levels for KIM1, TLR2, TLR4, IL‐1α, IL‐6 and TNFα in the kidney, respectively. The result is normalized to GAPDH. The data are presented as mean ± S.E. ( n = 4). *** P < 0.001, ** P < 0.001 versus sham group; ### P < 0.001, # P < 0.05 versus I/R group.

Article Snippet: Rat HMGB1 ELISA Kit was purchased from CUSABIO (Hubei, China).

Techniques: Control, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Quantitative RT-PCR, Expressing, Real-time Polymerase Chain Reaction

Journal: Nutrition & Metabolism

Article Title: Moderate doses of conjugated linoleic acid reduce fat gain, maintain insulin sensitivity without impairing inflammatory adipose tissue status in mice fed a high-fat diet

doi: 10.1186/1743-7075-7-5

Figure Lengend Snippet: Gene-specific primer sequences used in real-time PCR amplification

Article Snippet: Adiponectin and insulin plasma concentrations were measured using a rat/mouse adiponectin ELISA kit (Phoenix Europe GmbH, Karlsruhe, Germany) and Insulin Mouse Ultrasensitive ELISA kit (DRGInstruments GmbH, Marburg, Germany) respectively.

Techniques: Real-time Polymerase Chain Reaction, Sequencing

Journal: Nutrition & Metabolism

Article Title: Moderate doses of conjugated linoleic acid reduce fat gain, maintain insulin sensitivity without impairing inflammatory adipose tissue status in mice fed a high-fat diet

doi: 10.1186/1743-7075-7-5

Figure Lengend Snippet: Effects of CLA treatment on plasma concentration of metabolites in mice

Article Snippet: Adiponectin and insulin plasma concentrations were measured using a rat/mouse adiponectin ELISA kit (Phoenix Europe GmbH, Karlsruhe, Germany) and Insulin Mouse Ultrasensitive ELISA kit (DRGInstruments GmbH, Marburg, Germany) respectively.

Techniques: Clinical Proteomics, Concentration Assay

Journal: Nutrition & Metabolism

Article Title: Moderate doses of conjugated linoleic acid reduce fat gain, maintain insulin sensitivity without impairing inflammatory adipose tissue status in mice fed a high-fat diet

doi: 10.1186/1743-7075-7-5

Figure Lengend Snippet: Relative expression of target mRNAs in mature adipocytes and stromal vascular fraction in mice treated with CLA

Article Snippet: Adiponectin and insulin plasma concentrations were measured using a rat/mouse adiponectin ELISA kit (Phoenix Europe GmbH, Karlsruhe, Germany) and Insulin Mouse Ultrasensitive ELISA kit (DRGInstruments GmbH, Marburg, Germany) respectively.

Techniques: Expressing

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Repositioning Linagliptin for the Mitigation of Cadmium-Induced Testicular Dysfunction in Rats: Targeting HMGB1/TLR4/NLRP3 Axis and Autophagy.

doi: 10.3390/ph15070852

Figure Lengend Snippet: Figure 4. Effect of cadmium and/or linagliptin administration on the testicular pro-inflammatory response and HMGB1/TLR4/NF-κB pathway in rats. The co-treatment with linagliptin lowered tes- ticular TNF-α (A) and augmented IL-10 (B) alongside curtailing the testicular HMGB1/TLR4/NF-κB pathway that was stimulated by cadmium metal. The later event was demonstrated by downregulated protein expression of the high mobility group box protein 1 (HMGB1; (C)), toll-like receptor 4 (TLR4; (D)), and nuclear expression of the nuclear factor kappa B (NF-κBp65; (E)). * p < 0.05, ** p < 0.01, and *** p < 0.001 show statistical significance compared to the group of control rats; # p < 0.05, and ## p < 0.01 show statistical significance compared to the group of cadmium-intoxicated rats. Val- ues are stated as the mean ± standard error of the mean for 6 animals per group. LIN, linagliptin; Cd, cadmium chloride.

Article Snippet: Commercial ELISA kits were used for the measurement of TLR4 (Cusabio Technology, Houston, TX, USA; Catalog # CSB-E15822r), HMGB1 (Elabscience, Wuhan, China; Catalog # E-EL-R0505), and NF-κBp65 (Elabscience ELISA kit, Wuhan, China; Catalog # E-EL-R0674) in the testicular homogenate.

Techniques: Expressing, Control

Journal: Pharmaceuticals (Basel, Switzerland)

Article Title: Repositioning Linagliptin for the Mitigation of Cadmium-Induced Testicular Dysfunction in Rats: Targeting HMGB1/TLR4/NLRP3 Axis and Autophagy.

doi: 10.3390/ph15070852

Figure Lengend Snippet: Figure 11. An outline of the mechanisms that intercede linagliptin’s amelioration of cadmium-evoked testicular impairment. According to the current findings, the proposed mechanisms are: (A,A′) Curb- ing the inflammatory responses, including HMGB1/TLR4/NF-κB and NLRP3/caspase-1/IL-1β pathways in the testes of cadmium-intoxicated rats. (B) Stimulation of the autophagy response with activation of AMPK/mTOR pathway in the testes of cadmium-intoxicated rats. (C) Interference with the pro-apoptotic responses, including Bax downregulation, Bcl2 upregulation, and caspase 3 inhibition in the testes of cadmium-intoxicated rats. Of note, the beneficial outcomes of linagliptin are mainly due to DPP-4 inhibition, which has been linked to marked anti-inflammatory/immune- modulatory actions [21,22]. DPP-4 inhibition triggers several downstream targets, including the upregulated protein expression of SDF-1α, which prompts intensified testicular repair and regen- eration [21,33]. Moreover, the beheld activation of autophagy may suppress the pro-inflammatory responses/NLRP3 inflammasome activation [12,20] and limit the proapoptotic responses [12,17,19] in the testes of cadmium-intoxicated rats. Activation is represented by solid arrows, whereas inhibition is depicted by blunt arrows. The question marks denote unknown mechanisms.

Article Snippet: Commercial ELISA kits were used for the measurement of TLR4 (Cusabio Technology, Houston, TX, USA; Catalog # CSB-E15822r), HMGB1 (Elabscience, Wuhan, China; Catalog # E-EL-R0505), and NF-κBp65 (Elabscience ELISA kit, Wuhan, China; Catalog # E-EL-R0674) in the testicular homogenate.

Techniques: Activation Assay, Inhibition, Expressing