Fig 2: PLC-gamma-2 antibody screening by immunofluorescence.THP-1 WT and PLCG2 KO cells were labelled with a green or a far-red fluorescent dye, respectively. WT and KO cells were mixed and plated to a 1:1 ratio in a 96-well plate with optically clear flat-bottom. Cells were stained with the indicated PLC-gamma-2 antibodies and with the corresponding Alexa-fluor 555 coupled secondary antibody including DAPI. Acquisition of the blue (nucleus-DAPI), green (WT), red (antibody staining) and far-red (KO) channels was performed. Representative images of the merged blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Antibodies were tested at 1.0 μg/mL. Antibody dilution used: ab109267** at 1/1300, ab133522** at 1/600, MAB3716* at 1/500, NBP2-52536* at 1/1000, 3872 at 1/1000, 34264** at 1/102, 55512** at 1/50, GTX111178 at 1/100, GTX111293 at 1/700, MA5-35670** at 1/700 and MA5-38600* at 1/1000. Bars = 10 μm. *Monoclonal antibody, **Recombinant antibody.

Fig 3: PLC-gamma-2 antibody screening by Western Blot.Lysates of THP-1 (WT and PLCG2 KO) either treated (+) or not treated (-) with PMA were prepared and 10 μg of protein were processed for Western Blot with the indicated PLC-gamma-2 antibodies. The Ponceau stained transfers of each blot are presented to show equal loading of WT and KO lysates and protein transfer efficiency from the acrylamide gels to the nitrocellulose membrane. Antibody dilutions were chosen according to the recommendations of the antibody supplier. All antibodies were tested at a dilution of 1/500 except antibody 34264** was tested at 1/200. Predicted band size: 148 kDa. *Monoclonal antibody, **Recombinant antibody.