Journal: The Journal of Cell Biology

Article Title: APC is required for muscle stem cell proliferation and skeletal muscle tissue repair

doi: 10.1083/jcb.201501053

Figure Lengend Snippet: APC is required for adult skeletal muscle regeneration. (A) TM regimen and cardiotoxin (CTX) injury scheme for control and APC SC-KO mice. (B and C) Immunostaining for Pax7 (B) and e-MyHC (C) on TA cryosections 4 d after injury. (D–L) Analysis of TA muscles 14 d after injury reveals that tissue regeneration is defective in APC SC-KO muscles. (D) Whole sections of TA muscles immunostained for Laminin. Immunostaining for Tcf4 (F), type 1 Collagen (H), and Pax7 and Laminin (L) on TA cryosections is shown. Quantification of TA weight 14 d after injury normalized by uninjured TA weight (E), of the number of Tcf4+ cells/mm 2 (G), of relative Collagen+ area (I), and of sublaminar Pax7+ cells (normalized by Pax7+ cells of uninjured TA; J) is also shown. (K) GFP immunostaining on transverse sections of TAs isolated from Pax7 CreER ;Z/EG (Control-Z/EG) and Pax7 CreER APC lox/lox ;Z/EG (APC SC-KO-Z/EG) 14 d after injury demonstrates that APC-null satellite cells do not give rise to new myofibers, nor trans-differentiate in other cell types. Bars: (B, C, F, H, K, and L) 50 µm; (D) 200 µm. Nuclei are stained with Hoechst. Error bars indicate SEM. Inset panels show a 2× enlargement.

Article Snippet: Primary antibodies were as follows: mouse β-catenin (BD), goat Collagen Type I (SouthernBiotech), rabbit GFP (Life Technologies), rabbit Ki67 (Abcam), rabbit Laminin (Sigma-Aldrich), mouse MyoD (Dako), mouse embryonic-MyHC (Developmental Studies Hybridoma Bank), mouse Pax7 (DSHB), and rabbit Tcf4 (Cell Signaling Technology).

Techniques: Immunostaining, Isolation, Staining

Journal: Frontiers in cellular neuroscience

Article Title: Comparing Effects of Transforming Growth Factor β1 on Microglia From Rat and Mouse: Transcriptional Profiles and Potassium Channels.

doi: 10.3389/fncel.2018.00115

Figure Lengend Snippet: FIGURE 1 | Transforming growth factor β1 (TGFβ1) receptor in microglia after Intracerebral hemorrhage (ICH), and general effects of TGFβ1 on microglia in vitro. (A) Microglia/macrophages increase in the hematoma (H) after ICH and express TGFβR1. Representative confocal micrographs taken at 7 days in the damaged ipsilateral rat striatum (left) and the undamaged contralateral striatum (right). The inset is a higher magnification image of the boxed area (scale bar, 20 µm) showing microglia and infiltrating macrophages in the hematoma labeled with CD11b (OX-42 antibody; red) and TGFβR1 (green). Cell nuclei are labeled with 4’-6-diamidino-2-phenylindole (DAPI; blue). Scale bar for main images is 100 µm. (B) The morphology of isolated microglia (stained for CD11b, red, as in (A) was assessed using phalloidin to stain F-actin (green) and DAPI (blue) to label nuclei. Images are representative of results from six separate cell cultures for each species. Scale bar is 100 µm. (C) Microglia proliferation was determined 24 h after TGFβ1 treatment using the CyQUANTTM assay. Results are shown as fold-change with respect to untreated (control, CTL) microglia (blue dashed line) and expressed as mean ± SEM for 9–11 individual cell cultures. (D) Apoptosis was monitored using the TiterTACSTM colorimetric assay at 24 h after TGFβ1 treatment. Results are shown as mean absorbance at 450 nm (mean ± SEM, 8–9 individual cultures). The blue dashed line indicates values for the negative controls, and the red dashed line indicates the positive controls. Significant differences are shown between control and TGFβ1 treated cells (∗). One symbol of indicates p < 0.05; 2, p < 0.01; 3, p < 0.001.

Article Snippet: Microglia/macrophages were labeled with antibodies against CD11b (OX42, 1:200, Bio-Rad/AbD Serotec, RRID:SCR_013553; Cat# MCA275G, RRID:AB_321301) and TGFβR1 (1:500, Abcam, RRID:SCR_012931; Cat# ab31013, RRID:AB_778352).

Techniques: In Vitro, Labeling, Isolation, Staining, Control, Colorimetric Assay