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Table 1 List of reagents and antibodies used

From: Oxidative stress and inflammation cause auditory system damage via glial cell activation and dysregulated expression of gap junction proteins in an experimental model of styrene-induced oto/neurotoxicity

Reagent type

Designation

Source

Identifiers

Additional information

Use

Antibody

Anti-IBA-1(rabbit monoclonal)

Cell signaling

Cat. No. #17198

IF (1:100)

Marker of microglia/macrophages

Antibody

Anti-rabbit IgG

AlexaFluor 546 (rabbit polyclonal)

Thermo fisher scientific

Cat. No. #A-10040

IF (1:400)

Secondary antibody for immunofluorescence analyses

Antibody

Anti-rabbit IgG

AlexaFluor 488 (rabbit polyclonal)

Thermo fisher scientific

Cat.No. #A-21206

IF (1:400)

Secondary antibody for immunofluorescence analyses

Antibody

Anti-mouse IgG

AlexaFluor 546 (mouse polyclonal)

Thermo fisher scientific

Cat.No. #A10036

IF (1:400)

Secondary antibody for immunofluorescence analyses

Nuclear Counterstains

DAPI

Thermo fisher scientific

Cat.No. D1306

IF (0.5 mg/mL)

Marker of cell nuclei

Antibody

Anti-GFAP (mouse monoclonal)

Cell signaling

Cat. No. #3670

IF (1:300)

WB (1:1000)

Marker of glial fibrillary acidic protein (glial cells and astrocytes)

Antibody

Anti-COX-2 (rabbit monoclonal)

Cell signaling

Cat. No. #12282

IF (1:500)

WB (1:1000)

Marker of cyclooxygenase2 protein (inflammatory marker)

Antibody

Anti-CtBP2 (mouse)

BD biosciences

Cat. No. 612044

IF (1:1000)

Marker of C-terminal-binding protein 2 (pre-synaptic contact)

Antibody

Anti-NF200 (rabbit polyclonal)

Sigma-Aldrich

Cat. No. N4142

IF (1:80)

Marker of neurofilaments (afferent fibers)

Antibody

Anti-CD68 (mouse monoclonal)

Abcam

Cat. No. ab955

WB (1:1000)

IF (1:100)

Phagocytic markers involved in microglia and macrophages activation

Antibody

Anti-cleaved-caspase 3 (rabbit polyclonal)

Millipore

Cat. No. #AB3623

WB (1:100)

Marker of cleaved-caspase-3 protein (active form in apoptotic pathway)

Antibody

Anti-Caspase-3 (rabbit polyclonal)

Santa cruz biotechnology

Cat. No. sc-7272

WB (1:1000)

Marker of total caspase-3 protein

Antibody

Anti-NF-κB (rabbit monoclonal)

Cell signaling

Cat. No. #8242

WB (1:1000)

Transcription factors of the nuclear factor κB (NF-κB) (inflammatory marker)

Antibody

Anti-TNF-α (mouse monoclonal)

Santa cruz biotechnology

Cat. No. sc-52746

WB (1:1000)

Tumor Necrosis Factor α (inflammatory marker)

Antibody

Anti-IL-1β (rabbit polyclonal)

Santa cruz biotechnology

Cat. No. sc-7884

WB (1:1000)

Interleukin-1 β (IL-1β) (inflammatory marker)

Antibody

Anti-CXCR1 (rabbit polyclonal)

Thermofisher

Cat. No. #PA5-95749

WB (1:1000)

Marker of chemokine receptors protein

Antibody

Anti-iNOS (rabbit polyclonal)

Thermofisher

Cat. No. # PA1-036

WB (1:1000)

Marker of Inducible Nitric Oxide Synthase (iNOS)

Antibody

Anti-Cx43 (rabbit polyclonal)

Abcam

Cat. No. ab62252

WB (1:8000)

Marker of Connexin-43

Antibody

Anti-Cx26 (mouse monoclonal)

Thermofisher

Cat. No. # 13-8100

WB (1 µg/ml)

Marker of Connexin-26

Antibody

Anti-Cx30 (rabbit polyclonal)

Thermofisher

Cat. No. #71-2200

WB (1 µg/ml)

Marker of Connexin 30

Antibody

Anti-Panx1 (rabbit polyclonal)

Thermofisher

Cat. No. #487900

WB (1 mg/ml)

Marker of Pannexin 1

Antibody

Anti-Nitro tyrosine (rabbit)

Cell signaling

Cat. No. #06-284

WB (1:1000)

Marker of protein tyrosine nitration

Antibody

Anti-4HNE (rabbit polyclonal)

Alpha diagnostic international

Cat. No. #HNE11-S

WB (1:1000)

Detect the 4-Hydroxy-2-nonenal (HNE) a marker of lipid peroxidation

Antibody

Anti-GAPDH (mouse monoclonal)

Abcam

Cat. No. ab8245

WB (1:10,000)

Loading control

Antibody

Anti-β-tubulin

(mouse monoclonal)

Cell signaling

Cat. No. #2146

WB (1:5000)

Loading control

Antibody

Anti-phospho-AMPA Receptor 1 (GluA1, Ser845)

(rabbit polyclonal)

Cell signaling

Cat. No. #8084

WB (1:1000)

Receptor function, trafficking, and conductance

Antibody

Anti-AMPA Receptor 1 (GluA1) (rabbit polyclonal)

Cell signaling

Cat. No. #13185

WB (1:1000)

Total GluA1 levels