Figure 6. ANO2 channels expressed in the thalamus.

(a) RT–PCR data assessing ANO2 expression demonstrated that ANO2 was transcribed in the thalamus; β-actin was used as a reference transcript. (b) Western blotting data showed that ANO2 protein was expressed in the thalamus. ANO2-overexpressed cells displayed a broader ANO2 band related to various glycosylation levels, as described previously³⁵. (c) Immunostaining data indicated that ANO2 (green) co-localized with the neuronal marker NeuN (blue), and that ANO2 was located in the soma of TC neurons (scale bar, 20 μm). (d) Immunostaining for ANO2 (green) showed that AAV-Scr-infected TC neurons (red, upper) clearly expressed ANO2 in the soma, but those infected with AAV-shANO2 (red, lower) expressed ANO2 at a much lower level (scale bar, 50 μm). (e) Quantification of ANO2 expression in virus-infected TC neurons confirmed that gene silencing of ANO2 reduced the intensity of ANO2 expression in mCherry-positive neurons (AAV-Scr n=17, AAV-shANO2 n=16, ***P=0.0000015, t-test). Error bars represent mean±s.e.