Fig 2: CMA pathway and validation of LAMP2A immunohistochemical staining. (a) Overview of chaperone-mediated autophagy (CMA). ① Recognition and binding of HSC70 to the KFERQ-motif of the target protein. ② Translocation of the complex to the lysosome. ③ Binding of the target protein to LAMP2A at the lysosomal membrane. ④ Formation of a multimeric LAMP2A complex. ⑤ Translocation and degradation of the target protein. (b, c) Specificity of LAMP2A immunohistochemistry. SKBR3 cells were transduced with lentiviral vectors containing a LAMP2A cDNA (OE) construct, an empty vector control, or shRNAs targeting LAMP2A mRNA (shLAMP2A_1-2). After selection, cells were subjected to LAMP2A western blot analysis (b) and immunohistochemistry (c).

Fig 3: HSPA8, examples of immunohistochemical staining: (A): Adenocarcinoma, IRS 3 × 3 = 9; (B) Adenocarcinoma, IRS 2 × 4 = 8; (C): Squamous cell carcinoma, IRS 2 × 4 = 8; (D): Squamous cell carcinoma, IRS 0 × 4 = 0; Objective magnification: 10×, Scale bar: 200 µm.

Fig 4: Kaplan-Meier curves (overall survival and disease free survival) for expression of autophagy-related proteins: (a) OS, LAMP2A; (b) DFS, LAMP2A; (c) OS, HSC70; (d) DFS, HSC70; (e) OS, combined; (f) DFS, combined.

Fig 5: Examples of immunohistochemical staining: (a–h) LAMP2A ((a, e) IRS 3x4 = 12; (b, f) IRS 3x2 = 6; (c, g) IRS 1x4 = 4; (d, h) IRS 0x0 = 0); (i–p) HSC70 ((i, m) IRS 3x4 = 12; (j, n) IRS 3x3 = 9; (k, o) IRS 1x4 = 4; (l, p) IRS 0x0 = 0). Objective magnification: (a–d) 13x, (e–h) 40x, (i–l) 13x, and (m–p) 40x.