Anti rrm2

The construction of the microarray and the following immunohistochemistry were carried out as described previously by Shanghai Biochip Company.³⁹ Glioma and normal tissue sections from glioma patients were deparaffinized, repaired with citrate antigen, blocked, and treated with 5% goat serum for 15 minutes at room temperature. The tissue sections were stained by diluted primary anti‐CDKL3 (1:200; Bioss) or anti‐RRM2 (1:400; Abcam) at 4°C overnight. Tissue sections were then stained with DAB, counterstained with hematoxylin, and dehydrated with ethanol. Images were captured using an ImageScope v11 (Leica). All sections were scored by two independent pathologists in a blinded fashion. In particular, the intensity of positive‐staining cytoplasm or nucleus of CDKL3 was scored on a scale of 0‐3 (0, negative; 1, light brown; 2, medium brown; and 3, dark brown), which was multiplied by the corresponding value of positive percentage (1, less than 25%; 2, 25%‐50%; 3, 50%‐75%; and 4, more than 75%). The two scores of cellular cytoplasm and nucleus were added to estimate the expression level of CDKL3, which was divided into the high or low cut by the median total score 4.

The WB was performed using conventional protocols which are available elsewhere. The primary antibodies were anti-c-PARP (Abcam, #ab32064), anti-HMGB1 (Abcam, #ab18256), anti-GPX4 (Abcam, #ab125066), anti-GAPDH (CST, Boston, MA, USA, #5174), anti-HBA1 (Abcam, #ab191183), anti-PLIN4 (Abcam, #ab234752), anti-NNMT (Abcam, #ab223513), anti-STMN1 (Abcam, #ab52630), anti-RRM2 (Abcam, #ab209995), anti-CAPG (Abcam, #ab155688), anti-HIC1 (Abcam, #ab49326), anti-HNF4A (R&D Systems, PP-H6939-00 and Abcam, #ab181604), anti-KAT2B (Abcam, #ab12188), anti-Myc (CST, #2276) and anti-FLAG (CST, #8146).