Interleukin-13 Receptor in Psoriatic Keratinocytes: Overexpression of the mRNA and Underexpression of the Protein Juan C. Cancino-Díaz, Elba Reyes-Maldonado,

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Interleukin-13 Receptor in Psoriatic Keratinocytes: Overexpression of the mRNA and Underexpression of the Protein Juan C. Cancino-Díaz, Elba Reyes-Maldonado, Carmen A. Bañuelos-Pánuco, Luis Jiménez-Zamudio, Ethel García-Latorre, Mario E. Cancino-Díaz Journal of Investigative Dermatology Volume 119, Issue 5, Pages 1114-1120 (November 2002) DOI: 10.1046/j.1523-1747.2002.19509.x Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Differential expression of IL-13Rα1 mRNA in lesional and nonlesional skin from patients with psoriasis. Panel A: 6% PAGE stained with silver, of reverse transcription–PCR differential display analysis of nonlesional (NL) and lesional (L) skin from three patients with psoriasis. Reverse transcription–PCR differential display was developed with T11G and Ap4 primers. The arrows indicate the differential band which corresponded to mRNA to IL-13Rα1. Lane M contains a DNA size marker. Panels B and C: northern blot analysis with total RNA of nonlesional (NL) and lesional (L) skin from one patient with psoriasis. The cDNA of 580 bp obtained from the reverse transcription–PCR differential display was used as probe (32P-labeled) to detect the IL-13Rα1 mRNA (panel B). The paper was washed and incubated another one with 32P-labeled probe to β-actin, which was obtained by reverse transcription–PCR. Journal of Investigative Dermatology 2002 119, 1114-1120DOI: (10.1046/j.1523-1747.2002.19509.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Reverse transcription–PCR expression analysis of IL-13Rα1 mRNA and of IL-4Rα mRNA in skin samples of patients and controls. Panel A: reverse transcription–PCR for IL-13Rα1 in nonlesional (NL) and lesional (L) skin biopsies from 10 patients with psoriasis (lanes 1–10). Reverse transcription–PCR of IL-13Rα1 in lesional skin biopsies from patients with other dermatologic diseases such as: atopic dermatitis (lane 11), polymorphous light eruption (lane 12), pemphigus vulgaris (lane 13), seborrheic keratoses (lane 14), lichen planus (lane 15), and dermatofibroma (lane 16). M=DNA size marker, C1=Cos-7 cells (used as positive control of expression of IL-13Rα1 and IL-4Rα). Panel B: reverse transcription–PCR of IL-13Rα1 in skin biopsies from 10 healthy subjects (lanes 1–10). Reverse transcription–PCR of IL-13Rα1 in nonlesional (lane 11) and lesional (lane 12) skin biopsies from psoriatic patient 1. Reverse transcription–PCR of IL-13Rα1 in nondifferentiated (lane 13), differentiated keratinocytes (lane 14), differentiated keratinocytes treated with IL-13 (lane 15). Reverse transcription–PCR of IL-13Rα2 in differentiated keratinocytes nontreated (lane 16) and treated with IL-13 (lane 17). Panel C: reverse transcription–PCR of IL-4Rα in nonlesional (NL) and lesional (L) skin biopsies from 10 patients with psoriasis (lanes 1–10). C1=Cos-7 cells. Panel D: reverse transcription–PCR of IL-4Rα in skin biopsies from 10 healthy subjects (lanes 1–10). Lane M contains a DNA size marker. Journal of Investigative Dermatology 2002 119, 1114-1120DOI: (10.1046/j.1523-1747.2002.19509.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Comparison of the expression of IL-13Rα1 mRNA (panel A) and IL-4Rα mRNA (panel B) in nonlesional (NL) and lesional (L) skin from psoriatic patients, skin from healthy subjects, and skin from patients with other dermatologic diseases (ODD). The intensity of each band was measured in the Imager Ready Apparatus (Alpha Innotech Corporation) and normalized against that of β-actin. Bars indicate the mean of intensity. Variance analysis was done by the Duncan method. Journal of Investigative Dermatology 2002 119, 1114-1120DOI: (10.1046/j.1523-1747.2002.19509.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Immunohistochemical detection of IL-13Rα1 and IL-4Rα in nonlesional (NL) and lesional (L) skin biopsies from psoriatic patients and healthy controls (HC) and differentiated (DK) and nondifferentiated keratinocytes (NDK). MoAb IL-13Rα1 staining: panel A to panel H. Rabbit polyclonal antibody IL-4Rα staining: panel J to panel L. Negative control with irrelevant isotype-matched reagent: panel I. Journal of Investigative Dermatology 2002 119, 1114-1120DOI: (10.1046/j.1523-1747.2002.19509.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions