Journal: STAR Protocols

Article Title: Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation

doi: 10.1016/j.xpro.2025.103678

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-CNPase clone 11-5B (1:200) , Merck (Millipore) , Cat#MAB326; RRID: AB_2082608.

Techniques: Recombinant, Knock-Out, Software

Journal: Biomolecules & Therapeutics

Article Title: A 3D “ In Vitro ” Model to Study Hyaluronan Effect in Nasal Epithelial Cell Line Exposed to Double-Stranded RNA Poly(I:C)

doi: 10.4062/biomolther.2019.126

Figure Lengend Snippet: Effect of Poly(I:C) stimulation on Muc5AC and Muc5B mucin secretion in the Apical wash of ALI culture of RPMI 2650 cells. RPMI 2650 were grown in ALI culture for 21 days and stimulated with Poly(I:C) (0 to 50 µg/mL) for 5 days: (A) dose-response of Muc5AC (n=3), and (B) Muc5B (n=3) secretion. Effect of different MW of HA (HMW-HA, MMW-HA, and LMW-HA) on (C) Muc5AC (n=3), and (D) Muc5B secretion (n=3) in RPMI 2650 cells stimulated with Poly(I:C) (50 µg/mL). Muc5AC and Muc5B concentration was assessed in the apical wash samples by using ELISA method (as described in Materials and Methods). Data are expressed as mean ± SD of the concentration expressed as µg/mL. Statistical analysis was performed by ANOVA followed by Fisher’s PLSD correction. # p <0.05, ## p <0.01 and ### p <0.001 vs. untreated cells, and * p <0.05 *** p <0.001 vs. Poly(I:C) (50 mg/mL)-treated cells.

Article Snippet: After three washes, plates were incubated with a mouse monoclonal anti-Muc5AC Ab (45M1, cod SC-21701, Santa Cruz Biotechnology, Inc., CA, USA) or a mouse monoclonal anti-Muc5B Ab (5B#19-2E, cod SC-21768, Santa Cruz Biotechnology, Inc.).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Biomolecules & Therapeutics

Article Title: A 3D “ In Vitro ” Model to Study Hyaluronan Effect in Nasal Epithelial Cell Line Exposed to Double-Stranded RNA Poly(I:C)

doi: 10.4062/biomolther.2019.126

Figure Lengend Snippet: Effect of Poly(I:C) stimulation on Muc5AC and Muc5B mRNA expression in the cells from ALI culture of RPMI 2650 cells. RPMI 2650 were grown in ALI culture for 21 days and stimulated with Poly(I:C) for 5 days (A) dose-response of Poly(I:C) (0 to 50 µg/mL) on Muc5AC (n=3), and (B) Muc5B (n=3) mRNA. Effect of different MW of HA (HMW-HA, MMW-HA, and LMW-HA) on (C) Muc5AC (n=3), and (D) Muc5B mRNA (n=3) in RPMI 2650 cells stimulated with Poly(I:C) (50 µg/mL. Total RNA was extracted and Muc5B mRNA expression was assessed by real-time PCR (see the Materials and methods section for details). Relative quantitation of mRNA was carried out with the comparative CT method (2–ΔΔCt) and was plotted as fold-change compared to untreated cells chosen as the reference sample. Data are expressed as mean ± SD of fold change over untreated cells. Statistical analysis was performed by ANOVA followed by Fisher’s PLSD correction. # p <0.05 and ### p <0.001 vs. untreated cells, * p <0.05 vs. Poly(I:C) (50 mg/mL)-treated cells, ° p <0.05 and °°° p <0.01 vs. Poly(I:C) (50 mg/mL)+HHA-treated cells.

Article Snippet: After three washes, plates were incubated with a mouse monoclonal anti-Muc5AC Ab (45M1, cod SC-21701, Santa Cruz Biotechnology, Inc., CA, USA) or a mouse monoclonal anti-Muc5B Ab (5B#19-2E, cod SC-21768, Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitation Assay

Journal: Biomolecules & Therapeutics

Article Title: A 3D “ In Vitro ” Model to Study Hyaluronan Effect in Nasal Epithelial Cell Line Exposed to Double-Stranded RNA Poly(I:C)

doi: 10.4062/biomolther.2019.126

Figure Lengend Snippet: Graphical abstract: Effect of HMW-HA on TEER measurements, pH values, rheological properties, and Muc5AC and Muc5B production in an “ in vitro ” model of nasal viral infection.

Article Snippet: After three washes, plates were incubated with a mouse monoclonal anti-Muc5AC Ab (45M1, cod SC-21701, Santa Cruz Biotechnology, Inc., CA, USA) or a mouse monoclonal anti-Muc5B Ab (5B#19-2E, cod SC-21768, Santa Cruz Biotechnology, Inc.).

Techniques: In Vitro, Infection

Journal: eLife

Article Title: Switching of RNA splicing regulators in immature neuroblasts during adult neurogenesis

doi: 10.7554/eLife.87083

Figure Lengend Snippet: ( A ) Population doublings (PD). Data were obtained from three independent experiments (n=8). ( B ) Clonogenicity assay. Results were obtained from two independent experiments. The number inside the bars indicated the number of microplate wells analyszed (mean ± SEM). ( C ) Differentiation assay. Representative images of immunofluorescence of freshly sorted iNB cells cultured in oligodendrocyte, astrocyte, or neuronal differentiation medium, and stained for NG2 and CNPase, GFAP and CD133 or βIII-Tubulin and doublecortin (DCX) expressions, respectively. *p<0.05, **p<0.01, ***p<0.001 (Mann-Whitney U-test). Scale bar: 20 µm. ( D ) EGFP-positive s-iNB were isolated from β−actin:eGFP mice and transplanted unilaterally at three injection points at proximity of the dSVZ/RMS of recipient C57Bl/6 J mice. Transplanted brains were analyszed 5 weeks later by immunostaining. ( a ) Detection of eGFP + cells in the granule cell layer and the external cell layer of the olfactory bulb of a mouse transplanted. (a1–a2) a2 high magnification of the inset (dotted line in a) showing eGFP + NeuN + cells (white arrows). ( b–c ) Detection of eGFP + GFAP + DCX + cells (yellow arrows) and eGFP +GFAP + (white arrows) in the dorsal ( b2 ) and lateral ( c2 ) SVZ. GCL:granule cell layer, EPL: external plexiform layer, GL: glomerular layer, lSVZ: lateral SVZ, dSVZ: dorsal SVZ, Scale bars = 40 µm or 100 µm.

Article Snippet: Antibody , Mouse monoclonal anti-mouse CNPase (clone 11-5B) , Merck Millipore , Cat#MAB326 , 1:100.

Techniques: Differentiation Assay, Immunofluorescence, Cell Culture, Staining, MANN-WHITNEY, Isolation, Injection, Immunostaining

Journal: eLife

Article Title: Switching of RNA splicing regulators in immature neuroblasts during adult neurogenesis

doi: 10.7554/eLife.87083

Figure Lengend Snippet: Immunofluorescence of mouse brains transplanted unilaterally in 3 injection points near the dSVZ/RMS with eGFP + iNB cells isolated from β-actin:eGFP mice. Transplanted brains were analyszed 5 weeks later. ( A ) Example of EGFP + iNB cell (white arrow) that has reached the rostral migratory stream and expressed DCX (inset a3), but not GFAP (inset a2). Insets are high magnification panels. ( B ) eGFP + cells persisting at IS expressing astrocyte or oligodendrocyte markers. High magnifications of the inset1 (dotted line in A) showing eGFP + GFAP + cells (white arrow) (a2) and eGFP + DCX + cells (white arrow) (a3). High magnifications of the inset2 (dotted line in B) showing eGFP + CNPase + (yellow arrow) and eGFP + CNPase - (white arrow). Scale bars = 100 µm. ( C ) Quantitative analysis of GFAP, CNPase or DCX antibodies staining of GFP-positive cells persisting at IS (where high number of grafted cells were found) in ( A, B, C ). This was performed by using the NIS software measuring eGFP-, GFAP-, CNPase- and DCX-positive areas, n = 7-8 mice brains. The intersections between each marker and eGFP areas were determined as a percentage of staining. The results showed that approximately one third of eGFP + cells expressed GFAP or DCX. The quantitative analysis of CNPase expression specific to eGFP + cells was complicated by host cells, but an overexpression in eGFP-positive areas clearly revealed the expression of CNPase by a significant faction of these cells.

Article Snippet: Antibody , Mouse monoclonal anti-mouse CNPase (clone 11-5B) , Merck Millipore , Cat#MAB326 , 1:100.

Techniques: Immunofluorescence, Injection, Isolation, Expressing, Staining, Software, Marker, Over Expression

Journal: eLife

Article Title: Switching of RNA splicing regulators in immature neuroblasts during adult neurogenesis

doi: 10.7554/eLife.87083

Figure Lengend Snippet: ( A ) Clonogenic potential of FACS-isolated eGFP + iNB and eGFP - iNB, and recombined eGFP + mNB isolated from DCX-CreERT2::CAG-floxed-eGFP mice, 7 days after the first injection of tamoxifen. ( B ) Population doublings (PD) of FACS-isolated eGFP + iNB and eGFP - iNB, eGFP + mNB cells and total iNB cells (regardless of eGFP status). No statistical difference was found in the growth rate of the different isolated populationss, Mann-Whitney U-test., (n=8) ( C ) Representative images of immunofluorescence of freshly sorted eGFP + iNB cells cultured in oligodendrocyte,astrocyte, or neuronal differentiation medium, and stained for NG2 and CNPase, GFAP and CD133, βIII-Tubulin and DCX, respectively. Scale: 10 µm. ( D ) Quantification of the percentage of eGFP + iNB among DAPI cells cultured in differentiation media that differentiate in either neurons (βIII-tubulin), oligodendrocytes (NG2) or astrocytes (GFAP).

Article Snippet: Antibody , Mouse monoclonal anti-mouse CNPase (clone 11-5B) , Merck Millipore , Cat#MAB326 , 1:100.

Techniques: Isolation, Injection, MANN-WHITNEY, Immunofluorescence, Cell Culture, Staining

Journal: eLife

Article Title: Switching of RNA splicing regulators in immature neuroblasts during adult neurogenesis

doi: 10.7554/eLife.87083

Figure Lengend Snippet:

Article Snippet: Antibody , Mouse monoclonal anti-mouse CNPase (clone 11-5B) , Merck Millipore , Cat#MAB326 , 1:100.

Techniques: Recombinant, Red Blood Cell Lysis, Software, Staining, Immunolabeling, FACS

Journal: Dose-Response

Article Title: Comparative Analysis of Mitochondrial Proteome Reveals the Mechanism of Enhanced Ram Sperm Motility Induced by Carbon Ion Radiation After In Vitro Liquid Storage

doi: 10.1177/1559325818823998

Figure Lengend Snippet: Differential Mitochondrial Proteins in Ram Sperm Subjected to Carbon ion Radiation After 24 Hours of In Vitro Liquid Storage at 5°C.

Article Snippet: The 5 commercial primary antibodies used were rabbit polyclonal IgG anti-COX5B (Cat. #bs-3931R); mouse polyclonal IgG anti-ERAB/HSD17B10 (Cat. #bs-0021M); rabbit polyclonal IgG anti-ETFA (Cat. #bs-0494R); rabbit polyclonal IgG anti-SDHB (Cat. #bs-6650R); rabbit polyclonal IgG anti-SOD2 (Cat. #bs-20667R; Bioss Biotechnology Co, Ltd, Beijing, China), and rabbit polyclonal IgG anti-β-actin (Cat. #AP0060; Bioworld technology, Inc, Ltd, Nanjing, China).

Techniques: In Vitro

Journal: PLoS ONE

Article Title: T-Helper Type-2 Contact Hypersensitivity of Balb/c Mice Aggravated by Dibutyl Phthalate via Long-Term Dermal Exposure

doi: 10.1371/journal.pone.0087887

Figure Lengend Snippet: Representative images of the expression of (A) IL-5 and (B) IL-13 as determined by immunohistochemical staining (brown color stain) (black arrow). Panel: (A1, B1) Saline, (A2, B2) DBP 0.4, (A3, B3) DBP 4, (A4, B4) DBP 40, (A5, B5) FITC, (A6, B6) FITC+DBP 0.4, (A7, B7) FITC+DBP 4, (A8, B8) FITC+DBP 40. Analyses of (C) IL-5 and (D) IL-13 expression levels according to average optical density. Animal groups (in all panels): n = 4 mice per group. **: p <0.01, compared with saline group; ##: p <0.01, compared with FITC group.

Article Snippet: Mouse anti-IL-5-antibody, mouse anti-IL-13-antibody, and mouse anti-eosinophil cationic protein (ECP)-antibody, goat-anti-rabbit lgG-antibody, a rabbit lgG peroxidase conjugated streptavidin-biotin complex (SABC-POD) kit and a diaminobenzidine (DAB) kit were obtained from Boster Bio-engineering (Wuhan, China).

Techniques: Expressing, Immunohistochemical staining, Staining