Defective nuclear localization of Hsp70 in MDS erythroblasts. (A) Quantification of HSP701B transcript. RT-qPCR in 9 MDS () and 7 controls (□) erythroblasts, expressed as mean normalized relative quantity (n_RQ) ± SE to GAPDH. (B) Immunoblot for Hsp70 in erythroblasts. (Left panel) Whole-cell lysates. Hsc70 as loading control. Representative of 2 separated experiments. (Right panel) Total Hsp70 expression quantified by densitometry as a ratio to Hsc70 normalized to the ratio of Hsp70 to Hsc70 in UT-7 control cell line for each blot. Student t test for P values; *P < .05. (C) Subcellular localization of Hsp70 by immunofluorescence. MDS with (n = 33) or without dyserythropoiesis (n = 11) or control erythroblasts (n = 19) labeled with Ab to Hsp70. DAPI for nuclei. MGG-stained cytospins are shown (left panel). Nucleo/cytosolic (N/C) ratio of Hsp70 expression at day 14 using the ImageJ software (right panel; **P < .01). (D) Immunoblot analysis of Hsp70. Immunoblot for Hsp70 and Hsc 70 in nuclear and cytosolic extracts (left panel) in MDS with (n = 19) or without (n = 3) dyserythropoiesis and controls (n = 9). Quantification by densitometry of N/C ratio of Hsp70 normalized to N/C ratio of Hsc70 (means ± SE; right panel; *P < .05).