fluo 3 am  (Biotium)

Journal: NPJ Parkinson's Disease

Article Title: Loss of METTL14 in dopaminergic neurons disrupts ER homeostasis via m6A-dependent regulation of Atp2a3 mRNA: Implications for Parkinson’s Disease

doi: 10.1038/s41531-026-01318-7

Figure Lengend Snippet: A , B SH-SY5Y cells were transfected with an empty pCRIPERV2 vector (NC) or pCRIPERV2-gRNA plasmids (Mettl14-KD#1 and Mettl14-KD#2), 12 h later, cellular Ca 2+ was detected by FLUO-3 AM probe and flow cytometry. C SH-SY5Y cells were treated the same as ( A ), 12 h later, cellular Ca 2+ was observed under microscope (bar = 10 μm). D The endoplasmic reticulum and mitochondria of neurons in SN of Ctrl and Mettl14-cKO mice were observed under a transmission electron microscope (bar = 1 μm left or 500 nm right). E , F The distance between ER and outer mitochondrial membrane (OMM) and the number of ER-Mito contacts per 10 μm Mitochondrial membrane surface were analyzed ( n = 4). G SH-SY5Y cells were treated the same as ( A ), 12 or 24 h later, ER stress marker proteins GRP78, ATF4 and CHOP expression were detected by WB assay. H – J Quantification of WB. All data are presented as mean ± SD, ns indicated no significant, * P < 0.05, ** P < 0.01 compared to NC.

Article Snippet: Cell membrane permeable calcium fluorescent probe Fluo-3 AM (40703ES50) was purchased from Yeasen Biotechnology.

Techniques: Transfection, Plasmid Preparation, Flow Cytometry, Microscopy, Transmission Assay, Membrane, Marker, Expressing

Journal: NPJ Parkinson's Disease

Article Title: Loss of METTL14 in dopaminergic neurons disrupts ER homeostasis via m6A-dependent regulation of Atp2a3 mRNA: Implications for Parkinson’s Disease

doi: 10.1038/s41531-026-01318-7

Figure Lengend Snippet: A , B SH-SY5Y cells were transfected with pCRIPERV2-gRNA#1 plasmid (Mettl14-KD) or pCMV-Atp2a3-cDNA coding plasmid (Atp2a3-OE), combined with their empty vectors (Control: cells were transfected with empty pCRIPERV2 and empty pCMV vectors; Mettl14-KD: cells were transfected with pCRIPERV2-gRNA#1 and empty pCMV vector; KD&Atp2a3-OE: cells were transfected with pCRIPERV2-gRNA#1 and pCMV-Atp2a3-cDNA coding plasmid). Twelve hours later, cellular Ca 2+ was detected by FLUO-3 AM probe and flow cytometry. C SH-SY5Y cells were treated the same as ( A ), 12 h later, ER stress marker proteins GRP78, ATF4 and CHOP expression were detected by WB assay. D – F Quantification of WB. G , H SH-SY5Y cells were treated the same as ( A ), 12 h later, cells were treated DCFH-DA for 15 min and the cellular ROS was detected by flowcytometry. The mean DCF fluorescence intensity were quantified as relative DCFH-DA DCFH-DA level. I SH-SY5Y cells were treated the same as ( A ), 24 h later, relative cell number was quantified by cell counting assay. All data are presented as mean ± SD. * P < 0.05, ** P < 0.01 compared to Control; ## P < 0.01 compared to Mettl14-KD.

Article Snippet: Cell membrane permeable calcium fluorescent probe Fluo-3 AM (40703ES50) was purchased from Yeasen Biotechnology.

Techniques: Transfection, Plasmid Preparation, Control, Flow Cytometry, Marker, Expressing, Fluorescence, Cell Counting