Journal: Cell Reports Medicine

Article Title: Intratumoral erythroblastic islands restrain anti-tumor immunity in hepatoblastoma

doi: 10.1016/j.xcrm.2023.101044

Figure Lengend Snippet:

Article Snippet: The primary antibodies in the panel showing the interaction of erythroid cell with Macrophage (Panel 1) were: anti-GYPA (Abcam, Cat# ab129024, 1:10000), anti-CD68 (CST, Cat# 76437S, 1:1000), anti-VCAM (Abcam, Cat# ab134047, 1:500) and anti-CD45 (CST, Cat# 13917S, 1:500).

Techniques: Recombinant, Purification, Giemsa Stain, Red Blood Cell Lysis, Selection, Activation Assay, Expressing, Microarray, Software

Journal: Neural Regeneration Research

Article Title: In situ direct reprogramming of astrocytes to neurons via polypyrimidine tract-binding protein 1 knockdown in a mouse model of ischemic stroke

doi: 10.4103/1673-5374.390957

Figure Lengend Snippet: Vascular remodeling and reactive microglia in ischemic foci after in situ reprogramming. (A) Representative image of CD31 immunostaining (red, Alexa Fluor® 594, arrows) in ischemic foci at 60 dpi. The morphology of CD31-positive vasculature was compromised in the shCon group. CD31-positive vasculature in the shPTB group was noticeably improved. Scale bars: 100 μm. (B) Representative image of Iba1 immunostaining (red, Alexa Fluor® 594) in ischemic foci at 7, 14, 40, and 60 dpi. There were fewer Iba1-positive cells in the shPTB group compared with the shCon group. Scale bars: 100 μm. (C) Quantification of Iba1-positive area in B. Data are expressed as mean ± SD (7–40 dpi: n = 3; 60 dpi: n = 6). **** P < 0.0001 (one-way analysis of variance with Bonferroni post hoc test). (D) Representative image of CD68 immunostaining (red, Alexa Fluor® 594) in ischemic foci (AP: +0.2 mm) at 7, 14, 40, and 60 dpi. There were fewer CD68-positive cells in the shPTB group compared with the shCon group. Scale bars: 100 μm. (E) Quantification of CD68-positive area in D. Data are expressed as mean ± SD (7–40 dpi: n = 3; 60 dpi: n = 6). **** P < 0.0001 (one-way analysis of variance with Bonferroni post hoc test). AP: Anterior-posterior; dpi: days post-injection; GFP: green fluorescent protein; Iba1: ionized calcium-binding adapter molecule 1.

Article Snippet: The primary antibodies were as follows: rabbit anti-ionized calcium-binding adaptor molecule 1 (Iba1; 1:500, Abcam, Cambridge, UK, Cat# ab178846, RRID: AB_2636859), rabbit anti-GFAP (1:500, Abcam, Cat# ab7260, RRID: AB_305808), rabbit anti-neuronal nuclear protein (NeuN; 1:250, Abcam, Cat# ab177487, RRID: AB_2532109), rabbit anti-vesicular glutamate transporter 1 (VGLUT1; 1:200, Abcam, Cat# ab180188, RRID: AB_2868428), rabbit anti-COUP-TF-interacting protein-2 (Ctip2; 1:200, Abcam, Cat# ab28448, RRID: AB_1140055), rabbit anti-CD68 (1:500, Abcam, Cat# ab125212, RRID: AB_10975465), rabbit anti-microtubule-associated protein 2 (MAP2; 1:500, Abcam, Cat# ab32454, RRID: AB_776174), and rabbit anti-CD31 (1:50, Abcam, Cat# ab28364, RRID: AB_726362).

Techniques: In Situ, Immunostaining, Injection, Binding Assay

Journal: Journal of Neuroinflammation

Article Title: The alternatively spliced fibronectin CS1 isoform regulates IL-17A levels and mechanical allodynia after peripheral nerve injury

doi: 10.1186/s12974-015-0377-6

Figure Lengend Snippet: FN in CCI nerve. a Fibronectin (FN) exists as a heterodimer linked by disulfide bonds. One of the monomers includes the 25-amino-acid-long CS1 sequence. Typical CS1 inclusion and exclusion splice variants are illustrated. b FN immunoblot of rat sciatic nerve, normal or at days 1 and 7 after CCI. β-actin, a loading control (40 μg total protein). c The graph represents the mean levels of the individual FN isoform recorded in ( b ). ±SEM relative to β-actin ( n = 3–4/group; * p < 0.05, ** p < 0.01, n.s. not significant). d FN-CS1 immunostaining ( red ) in sciatic nerves at day 7 post-CCI. The vessel ( V ) endothelial cells, Schwann cells, and macrophages (confirmed using the CD68 immunostaining, green ) are stained. DAPI, blue . Representative of n = 3/group; scale bars, 40 μm. e FN-CS1 immunostaining ( red ) in a teased nerve fiber at day 7 post-CCI; representative of ~20 individual fibers from n = 3. Scale bar, 10 μm. NoR node of Ranvier

Article Snippet: For dual immunofluorescence, following the washes in PBS-1 % Tween, the slides were incubated for 1–2 h at ambient temperature with rabbit polyclonal CD68 antibody (Santa Cruz, cat. #SC9139), followed by incubation with goat anti-rabbit Alexa 488-conjugated secondary antibody (green, 1–2 h, ambient temperature).

Techniques: Sequencing, Western Blot, Control, Immunostaining, Staining

Journal: iScience

Article Title: An inducible explant model of osteoclast-osteoprogenitor coordination in exacerbated osteoclastogenesis

doi: 10.1016/j.isci.2023.106470

Figure Lengend Snippet: Key resources table

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-snx10 antibody Sigma Cat#sab2107086 Mouse monoclonal anti-tgs101 antibody Novus Cat#nb200-112 Rat monoclonal anti-CD68 antibody Abcam Cat#ab53444 Mouse monoclonal anti-runx2 antibody Abcam Cat#ab76956 Rabbit monoclonal anti-ki67 antibody Cell signaling technologies Cat#ab76956 Mouse monoclonal anti-rank antibody Abcam Cat#ab13918 Rabbit polyclonal anti-ctsk antibody Invitrogen Cat#pa5-102483 Rabbit polyclonal anti-gnas antibody Millipore Cat#371732 Rabbit monoclonal anti-cyclophilin b antibody Cell signaling technologies Cat#43603s Goat polyclonal anti-mouse secondary antibody Cell signaling technologies Cat#4408 Goat polyclonal anti-mouse secondary antibody Cell signaling technologies Cat#4409 Goat polyclonal anti-rabbit secondary antibody Cell signaling technologies Cat#4412 Goat polyclonal anti-rabbit secondary antibody Cell signaling technologies Cat#4413 Goat polyclonal anti-rat secondary antibody Invitrogen Cat#a-1106 Goat polyclonal anti-mouse secondary antibody Abcam Cat#7069 Goat polyclonal anti-rabbit secondary antibody Abcam Cat#7091 Biological samples Murine bone marrow isolates Collins laboratory N/A Chemicals, peptides, and recombinant proteins Doxycycline hyclate Sigma Cat#d9891 cOmplete protease inhibitor Sigma Cat#118361700010 Normocin Invivogen Cat#ant-nr-1 ECF substrate Cytiva Cat# rpn5785 Hoechst 33342 Invitrogen Cat#h3570 Phalloidin-Alexa488 Invitrogen Cat#a30106 Recombinant mouse osteoprotegerin (fc tag) Cell Sciences Cat#crm761b-fc Critical commercial assays Bone demineralization assay CosmoBio Cat#csr-bra-24kit TRAP staining assay CosmoBio Cat#pmc-ak04f-cos Murine m -csf elisa assay Abcam Cat#ab199084 Murine rankl elisa assay Abcam Cat#ab100749 Murine il-6 elisa assay Abcam Cat#ab222503 Murine lif elisa assay Abcam Cat#ab238261 Beta-galactosidase staining assay Sigma Cat#gals-1kt PureLink RNA extraction kit Invitrogen Cat#12183018a iQ SYBR Green Supermix Biorad Cat#1708882 iScript RNA-to-cDNA kit Biorad Cat#1708841 Experimental models: Cell lines MC3T3-E1 clone 4 ATCC Cat#crl-2593 MC3T3-E1 clone 14 ATCC Cat#crl-2594 Experimental models: Organisms/strains Prrx1-Cre/LSL-rtTA-IRES-GFP/TetO-GαsR201C/TetO-βgal mice Collin’s Laboratory NICHD, NIH N/A Oligonucleotides Nfactc1 primers Sigma PrimerPairID# M1_Nfatc1 Fos primers Sigma PrimerPairID# M1_Fos Sp7 primers Sigma PrimerPairID# M1_Sp7 Dlx5 primers Sigma PrimerPairID# M1_Dxl5 Bglap primers Sigma PrimerPairID# M1_Bglap 18s rRNA primers Sigma PrimerPairID# M1_Rn18s Actin primers Sigma PrimerPairID# M1_Actb Open in a separate window Key resources table .

Techniques: Recombinant, Protease Inhibitor, Staining, Enzyme-linked Immunosorbent Assay, RNA Extraction, SYBR Green Assay