Journal: Developmental cell
Article Title: CRIPTO-based micro-heterogeneity of mouse muscle satellite cells enables adaptive response to regenerative microenvironment.
doi: 10.1016/j.devcel.2023.11.009
Figure Lengend Snippet: Figure 4. Regenerative potential of CRIPTONeg and CRIPTOPos cells and reconstitution of naı¨ve CRIPTO micro-heterogeneity following in vivo cell transplantation (A) Schematic representation of the experimental design. (B) Representative images of mosaic recipient muscles showing tdTomato-positive myofibers (RFP-red) and double staining with GFP (green) and LAMININ (white). (C and D) Quantification of tdTomato+ myofibers number/area (C) and GFP+ cells from CRIPTONeg and CRIPTOPos donor cells normalized per mm2 (D). Scale bars: 100 mm. Data are mean ± SEM. Nuclei were counter-stained with DAPI. (n = 3). (E) Schematic representation of the experimental design. (F and G) Representative dot plots of surface CRIPTO in GFP-positive CRIPTONeg and CRIPTOPos cells used for transplantation (F) and GFP-positive ASCs (3 dpi) derived from CRIPTONeg and CRIPTOPos -transplanted mice (G). The percentage of CRIPTO-positive cells above the threshold is indicated in the dot plots.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse Cripto Allophycocyanin MAb Biotechne - R&D systems Cat# FAB1538A Cripto Mouse specific Cell Signaling Technology Cat# 2818; RRID: AB_2240508 Anti-P4HB antibody [RL90] abcam Cat# ab2792; RRID: AB_303304 Pax7 Antibody Developmental Studies Hybridoma Bank (DHSB) Cat# PAX7; RRID: AB_528428 Anti-MyoD (C-20) Santa Cruz Biotechnology, Inc Cat# sc-304; RRID: AB_631992 Anti-GFP antibody abcam Cat# ab13970; RRID: AB_300798 Anti-human Ki67 Monosan Cat# MONX10283 Myogenin Antibody (F5D) Developmental Studies Hybridoma Bank (DHSB) Cat# F5D, RRID: AB_2146602 BrdU antibody [BU1/75 (ICR1)] GeneTex Cat# GTX26326; RRID: AB_1081056 Ki-67 Recombinant Rabbit Monoclonal Antibody (SP6) ThermoFisher Scientific Cat# MA5-14520; RRID: AB_10979488 Anti-Laminin Sigma-Aldrich L9393; RRID: AB_477163 Biotin-SP (long spacer) AffiniPure Goat Anti-Mouse IgG, Fcg subclass 1 specific Jackson ImmunoResearch Cat# 115-065-205, RRID: AB_2338571 Anti-mouse Alexa Fluor 549 ThermoFisher Scientific Cat# A-21203; RRID: AB_2535789 Anti-rabbit Alexa Fluor 488 ThermoFisher Scientific Cat# A-11001; RRID: AB_2534069 Anti-rabbit Alexa Fluor 555 ThermoFisher Scientific Cat# A-31572; RRID: AB_162543 Anti-rabbit Alexa Fluor 549 ThermoFisher Scientific Cat# A-21207; RRID: AB_141637 Alexa Fluor 488 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) ThermoFisher Scientific Cat# 703-545-155, RRID: AB_2340375 Cy 3 AffiniPure Donkey Anti-Rat IgG (H+L) Jackson ImmunoResearch Cat# 712-165-153, RRID: AB_2340667 Chemicals, peptides, and recombinant proteins Sambucus Nigra Lectin (SNA, EBL), Biotinylated Vector Laboratories Cat# B-1305-2 Rhodamine Phalloidin ThermoFisher Scientific Cat# R415 Streptavidin-Cy3 conjugated Jackson ImmunoResearch Cat# 016-160-084 Streptavidin-488 conjugated Invitrogen Cat# S32354 Streptavidin-647 conjugated Invitrogen Cat# 532357 Tamoxifene Sigma-Aldrich Cat# T5648 Cardiotoxin Latoxan Laboratory Cat# L8102 Dispase II Roche Cat# 10887800 Collagenase A Roche Cat# 10103586001 DNAse I Roche Cat# 11284932001 Hank’s Balanced Salt Solution (HBSS) Gibco Cat# 14170-088 Bovine Serum Albumin (BSA) Sigma-Aldrich Cat# A4503 Penicillin-Streptomycin Gibco Cat# 15140-122 Collagenase Sigma-Aldrich Cat# C0130 Dulbecco’s Modified Eagle’s medium (DMEM) Gibco Cat# 41965-039 Ham’s F-12 Nutrient Mix, GlutaMAX Gibco Cat# 31765-027 Pyruvate Sodium Gibco Cat# 11360-039 L-Glutamine 200mM (100x) Gibco Cat# 25030-024 (Continued on next page) e1 Developmental Cell 58, 2896–2913.e1–e6, December 18, 2023
Techniques: In Vivo, Transplantation Assay, Muscles, Double Staining, Staining, Derivative Assay
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