Journal: Cell stem cell

Article Title: Transient maternal IL-6 mediates long-lasting changes in neural stem cell pools by deregulating an endogenous self-renewal pathway.

doi: 10.1016/j.stem.2013.10.002

Figure Lengend Snippet: Figure 4. Embryonic Forebrain Precursors Express IL6R and Proliferate in Response to IL-6 (A) RT-PCR analysis for IL6R and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNAs in RNA from the E13 cortex (Ctx), from cortical precursors cultured for 1 day (CP culture), and from sorted E13 Sox2:EGFP-positive cortical precursors (sorted cells). RT denotes samples with (+) or without () reverse transcriptase.

Article Snippet: Validation of siRNA-Mediated Knockdown HEK293 cells were cultured and transfected as previously described (Vessey et al., 2012) with a murine FLAG-tagged IL6R plasmid (Origene) and either a control scrambled siRNA or a proprietary ‘‘Smartpool’’ IL6R siRNA cocktail (both from Thermo Scientific) for 24 hr.

Techniques: Reverse Transcription Polymerase Chain Reaction, Cell Culture, Reverse Transcription

Journal: Cell stem cell

Article Title: Transient maternal IL-6 mediates long-lasting changes in neural stem cell pools by deregulating an endogenous self-renewal pathway.

doi: 10.1016/j.stem.2013.10.002

Figure Lengend Snippet: Figure 7. Endogenous IL-6 and IL6Rs Regulate Embryonic Cortical Precursor Maintenance and Numbers In Vivo (A) Western blot of lysates from HEK293 cells cotransfected with a FLAG-tagged IL6R plasmid and scrambled or IL6R siRNAs, probed with a FLAG antibody. The blot was reprobed for ERK as a loading control. (B–G) E13/14 cortices were electroporated with an EGFP expression plasmid and IL6R or scrambled siRNAs, and cortices were analyzed 3 days later. (B and C) Coronal cortical sections were immunostained for EGFP (green, B) and the proportion of EGFP-positive cells in the different cortical regions was quantified (C). **p < 0.01; n = 3 each, 3–4 sections per embryo. In (B) the dotted lines demarcate the different cortical regions. (D–G) Cortical sections were immunostained for EGFP (green, D and F) and Sox2 (red, D) or Satb2 (red, F), and the proportion of EGFP-positive cells that were also positive for Sox2 (E) or Satb2 (G) was quantified. **p < 0.01; n = 3 each, 3–4 sections per embryo. Arrows in (D and F) indicate double-labeled cells. (H–M) Pregnant IL-6+/+ (WT) or IL-6/ mice were injected with BrdU at G13.5 and the cortices of their embryonic offspring were analyzed 3 days later. (H and I) Coronal cortical sections were immunostained for BrdU (red, H) and the proportion of BrdU-positive cells in different cortical regions was quantified (I). *p < 0.05; n = 5 embryos each. In (H) the dotted lines demarcate the different cortical regions.

Article Snippet: Validation of siRNA-Mediated Knockdown HEK293 cells were cultured and transfected as previously described (Vessey et al., 2012) with a murine FLAG-tagged IL6R plasmid (Origene) and either a control scrambled siRNA or a proprietary ‘‘Smartpool’’ IL6R siRNA cocktail (both from Thermo Scientific) for 24 hr.

Techniques: In Vivo, Western Blot, Plasmid Preparation, Control, Expressing, Labeling, Injection

Journal: BioMed Research International

Article Title: The Role of IL-6RA in UHMWPE Promotes Proliferation in Fibro-Like Synovial Cells

doi: 10.1155/2018/3928915

Figure Lengend Snippet: UHMWPE increased the expression of mIL-6R protein. FLS cells were incubated with UHMWPE (0, 0.01, 0.1, and 1g/L) for 7 d. The expression of IL-6R protein was detected by western blotting assay.

Article Snippet: The primary antibodies included caspase-3(Abcam, USA; ab4051), cleaved-caspase-3 (Cell Signaling Technology, USA; #9661), Bax (Abcam, USA; ab182733), Bcl-2 (Abcam, USA; ab692), IL-6R (Boster, China; A01425-1), β -actin (Bioss, China; bs-0061R), and GAPDH (Abcam, USA; ab8245) at 1:500 dilutions.

Techniques: Expressing, Incubation, Western Blot