primary mouse embryonic cf 1 fibroblasts  (ATCC)

Journal: Scientific Reports

Article Title: Male meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in mice

doi: 10.1038/s41598-024-55376-z

Figure Lengend Snippet: Unlike Mouse CF-1 Fibroblast Cells, TACC3 and cKAP5/chTOG immunolabel only mouse spermatogenic meiotic spindle poles but do not immunostain pre-prophase, interkinesis or post-meiotic spermatids including testicular sperm . Panel 1 : Metaphase GFP CETN2 CF-1 mouse fibroblast ( A , B : green) immunolabeled with γ-tubulin ( A , C : red), anti-TACC3 ( B , C : blue) and DNA ( A : inset, blue). TACC3 immunolabels the entire spindle including spindle kinetochore microtubules ( B , C , arrows). Insets: details; spindle pole TACC3 (blue), GFP-CETN2 centrioles ( B ; green) and γ-tubulin ( C : red). Panel 2 : Meiosis I ( A – E ) and meiosis-II ( A’ – E’ ) GFP CETN2-expressing spermatocytes (green), microtubules (red) and TACC3 (blue). TACC3 concentrates at metaphase spindle poles ( C , C’ : blue) but not kinetochore microtubules ( C – C’ : red, arrows). Upper insets: spindle pole TACC3 (blue) and GFP CETN2 centrioles (green); lower insets: GFP CETN2 (green), microtubules (red) and DNA (blue), except D’ : left, middle insets: microtubules, red; right inset: GFP CETN2 centrioles (green) and DNA (blue). Panel 3 : Metaphase GFP CETN2 CF-1 mouse fibroblast ( A , B : green) immunolabeled with γ-tubulin ( A , C : red), anti-cKAP5/chTOG ( B , C : blue) and DNA ( A : inset; blue). cKAP5/chTOG immunolabels the entire spindle including kinetochore microtubules ( B , C , arrows). Insets ( B , C ): details, spindle pole cKAP5/chTOG (blue), centrioles ( B : green) and γ-tubulin ( C : red). Panel 4: Meiosis-I ( A , B ) and meiosis-II ( A’ – B’ ) GFP CETN2-expressing spermatocytes (green), microtubules (red) and cKAP5/chTOG (blue). cKAP5/chTOG immunolabels spindle poles, not spindle kinetochore microtubules ( B : arrows). Upper insets: spindle pole GFP CETN2 centrioles (green) and cKAP5/chTOG (blue); lower insets: GFP CETN2 (green), microtubules (red) and DNA (blue). Panel 5 : Graph, right: analysis of > 60 TACC3 or cKAP5/chTOG meiotic spermatocytes showing similar TACC3 or cKAP5/chTOG spindle pole localization (ns, not significant; p < 0.5). All scale bars = 5 µm.

Article Snippet: Commercially available primary mouse embryonic fibroblasts isolated from CF-1 mice (Millipore; catalog # PMEF-CF) were thawed and propagated using protocols according to the manufacturer.

Techniques: Immunolabeling, Expressing

Journal: Scientific Reports

Article Title: Male meiotic spindle poles are stabilized by TACC3 and cKAP5/chTOG differently from female meiotic or somatic mitotic spindles in mice

doi: 10.1038/s41598-024-55376-z

Figure Lengend Snippet: TACC3 is restricted to spindle poles in male meiosis spermatocytes as opposed to the spindle LISD in female oocytes or spindle pole/kinetochore microtubules in fibroblast cells. Panel 1: A metaphase-I mouse GFP CETN2-expressing spermatocyte ( A : green, arrows) immunolabeled with γ-tubulin (A: red) and DNA (A: blue). Inset: TACC3 spindle pole immunolabeling (blue). ( B – D ) (upper pole) and ( B’ – D’ ) (lower pole) details; GFP CETN2 centrioles ( B , B’ : green), γ-tubulin ( C , C’ : red) and TACC3 ( D , D’ : blue). ( E , E’ ): overlays. Panel 2 : Metaphase-I mouse oocyte expressing GFP CETN2 ( A : green, arrows) at the spindle poles immunolabeled with pericentrin ( A : red) and DNA ( A : blue). Inset: spindle LISD TACC3 (blue). ( B – D ) (upper pole) and ( B’ – D’ ) (lower pole) details; GFP CETN2 foci ( B : green), spindle pole pericentrin MTOC ( C : red), and the spindle LISD TACC3 ( D : blue). ( E and E’ ) Overlays. Panel 3 : a mouse GFP CETN2-expressing CF-1 fibroblast cell ( A : green, arrows) immunolabeled with γ-tubulin ( A : red) and DNA ( A : blue). Inset: TACC3 immunolabels the spindle poles and kinetochore microtubules (blue, arrowheads). ( B – D ) (upper pole) and ( B’ – D’ ) (lower pole) details; GFP CETN2 centrioles ( B : green), γ-tubulin ( C : red) and TACC3 ( D : blue). ( E , E’ ) Overlays. Panels 1A, 2A, and 3A scale bars = 5 µm; Panels 1, 2, 3 ( B – E ) and ( B’ – E’ ) scale bars: 1 µm.

Article Snippet: Commercially available primary mouse embryonic fibroblasts isolated from CF-1 mice (Millipore; catalog # PMEF-CF) were thawed and propagated using protocols according to the manufacturer.

Techniques: Expressing, Immunolabeling